Solid forms of (1s,4s)-4-(2-(((3s,4r)-3-fluorotetrahydro-2h-pyran-4-yl)amino)-8-((2,4,6-trichlorophenyl)amino)-9h-purin-9-yl)-1-methylcyclohexane-1-carboxamide and methods of their use

ABSTRACT

Provided herein are formulations, solid forms and methods of use relating to (1s,4s)-4-(2-(((3S,4R)-3-fluorotetrahydro-2H-pyran-4-yl)amino)-8-((2,4, 6-trichlorophenyl)amino)-9H-purin-9-yl)-1-methylcyclohexane-1-carboxamide.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationNo. 62/317,468, filed Apr. 1, 2016, which is incorporated herein byreference in its entirety and for all purposes.

FIELD

Provided herein are solid forms ofcis-4-[2-{[(3S,4R)-3-fluorooxan-4-yl]amino}-8-(2,4,6-trichloroanilino)-9H-purin-9-yl]-1-methylcyclohexane-1-carboxamide,alternatively named(1s,4s)-4-(2-(((3S,4R)-3-fluorotetrahydro-2H-pyran-4-yl)amino)-8-((2,4,6-trichlorophenyl)amino)-9H-purin-9-yl)-1-methylcyclohexane-1-carboxamide,and methods of their use for the treatment of cancer.

BACKGROUND

The identification and selection of a solid form of a pharmaceuticalcompound are complex, given that a change in solid form may affect avariety of physical and chemical properties, which may provide benefitsor drawbacks in processing, formulation, stability, bioavailability,storage, handling (e.g., shipping), among other important pharmaceuticalcharacteristics. Useful pharmaceutical solids include crystalline solidsand amorphous solids, depending on the product and its mode ofadministration. Amorphous solids are characterized by a lack oflong-range structural order, whereas crystalline solids arecharacterized by structural periodicity. The desired class ofpharmaceutical solid depends upon the specific application; amorphoussolids are sometimes selected on the basis of, e.g., an enhanceddissolution profile, while crystalline solids may be desirable forproperties such as, e.g., physical or chemical stability (see, e.g., S.R. Vippagunta et al., Adv. Drug. Deliv. Rev., (2001) 48:3-26; L. Yu,Adv. Drug. Deliv. Rev., (2001) 48:27-42).

Whether crystalline or amorphous, solid forms of a pharmaceuticalcompound include single-component and multiple-component solids.Single-component solids consist essentially of the pharmaceuticalcompound or active ingredient in the absence of other compounds. Varietyamong single-component crystalline materials may potentially arise fromthe phenomenon of polymorphism, wherein multiple three-dimensionalarrangements exist for a particular pharmaceutical compound (see, e.g.,S. R. Byrn et al., Solid State Chemistry of Drugs, (1999) SSCI, WestLafayette). The importance of discovering polymorphs was underscored bythe case of Ritonavir™, an HIV protease inhibitor that was formulated assoft gelatin capsules. About two years after the product was launched,the unanticipated precipitation of a new, less soluble polymorph in theformulation necessitated the withdrawal of the product from the marketuntil a more consistent formulation could be developed (see S. R.Chemburkar et al., Org. Process Res. Dev., (2000) 4:413-417).

Notably, it is not possible to predict a priori if crystalline forms ofa compound even exist, let alone how to successfully prepare them (see,e.g., Braga and Grepioni, 2005, “Making crystals from crystals: a greenroute to crystal engineering and polymorphism,” Chem. Commun.: 3635-3645(with respect to crystal engineering, if instructions are not veryprecise and/or if other external factors affect the process, the resultcan be unpredictable); Jones et al., 2006, Pharmaceutical Cocrystals: AnEmerging Approach to Physical Property Enhancement,” MRS Bulletin31:875-879 (At present it is not generally possible to computationallypredict the number of observable polymorphs of even the simplestmolecules); Price, 2004, “The computational prediction of pharmaceuticalcrystal structures and polymorphism,” Advanced Drug Delivery Reviews56:301-319 (“Price”); and Bernstein, 2004, “Crystal Structure Predictionand Polymorphism,” ACA Transactions 39:14-23 (a great deal still needsto be learned and done before one can state with any degree ofconfidence the ability to predict a crystal structure, much lesspolymorphic forms)).

The variety of possible solid forms creates potential diversity inphysical and chemical properties for a given pharmaceutical compound.The discovery and selection of solid forms are of great importance inthe development of an effective, stable and marketable pharmaceuticalproduct.

The connection between abnormal protein phosphorylation and the cause orconsequence of diseases has been known for over 20 years. Accordingly,protein kinases have become a very important group of drug targets. (SeeCohen, Nature, 1:309-315 (2002), Gaestel et al. Curr. Med. Chem. 14:2214-223 (2007); Grimminger et al. Nat. Rev. Drug Disc. 9(12):956-970(2010)). Various protein kinase inhibitors have been used clinically inthe treatment of a wide variety of diseases, such as cancer and chronicinflammatory diseases, including rheumatoid arthritis and psoriasis.(See Cohen, Eur. J. Biochem., 268:5001-5010 (2001); Protein KinaseInhibitors for the Treatment of Disease: The Promise and the Problems,Handbook of Experimental Pharmacology, Springer Berlin Heidelberg, 167(2005)).

Cancer is characterized primarily by an increase in the number ofabnormal cells derived from a given normal tissue, invasion of adjacenttissues by these abnormal cells, or lymphatic or blood-borne spread ofmalignant cells to regional lymph nodes and to distant sites(metastasis). Clinical data and molecular biologic studies indicate thatcancer is a multistep process that begins with minor preneoplasticchanges, which may under certain conditions progress to neoplasia. Theneoplastic lesion may evolve clonally and develop an increasing capacityfor invasion, growth, metastasis, and heterogeneity, especially underconditions in which the neoplastic cells escape the host's immunesurveillance (Roitt, I., Brostoff, J and Kale, D., Immunology,17.1-17.12 (3rd ed., Mosby, St. Louis, Mo., 1993)).

Cancers figure among the leading causes of death worldwide, accountingfor 8.2 million deaths in 2012. It is expected that annual cancer caseswill rise from 14 million in 2012 to 22 million within the next twodecades (See Cancer Fact sheet No. 297, World Health Organization,February 2014, retrieved 10 Jun. 2014 and Globocan 2012, IARC).

The current drugs used in cancer treatment are highly toxic and oftennon-specific. Current anticancer therapy strategies are typicallyfocused on rapid proliferating cells, which can shrink primary andmetastatic tumors, but such effects are usually transient and tumorrelapse of most metastatic cancers frequently occur. One possible reasonfor failure is the existence of cancer stem cells. Unlike most cellswithin the tumor, cancer stem cells are resistant to well-definedchemotherapy, and after treatment, they can regenerate all the celltypes in the tumor through their stem cell-like behavior of largelyquiescent nature and their abundant expression of drug transporters.

There is an enormous variety of cancers which are described in detail inthe medical literature. The incidence of cancer continues to climb asthe general population ages, as new cancers develop, and as susceptiblepopulations (e.g., people infected with AIDS or excessively exposed tosunlight) grow. However, options for the treatment of cancer arelimited. A tremendous demand therefore exists for new methods andcompositions that can be used to treat patients with cancer.

Citation or identification of any reference in Section of thisapplication is not to be construed as an admission that the reference isprior art to the present application.

Accordingly, there remains a need for cancer therapies, for example,modulators, and in particular solid forms.

SUMMARY

Provided herein are solid forms of Compound 1:

having the name cis-4-[2-[(3S,4R)-3-fluorooxan-4-yl]amino-8-(2,4,6-trichloroanilino)-9H-purin-9-yl]-1-methylcyclohexane-1-carboxamide,alternatively named (1 s,4s)-4-(2-(((3S,4R)-3-fluorotetrahydro-2H-pyran-4-yl)amino)-8-((2,4,6-trichlorophenyl)amino)-9H-purin-9-yl)-1-methylcyclohexane-1-carboxamide,including tautomers thereof.

Also provided are methods of preparing, isolating, and characterizingthe solid forms.

In certain aspects, the solid forms of Compound 1 described herein areuseful for treating or preventing one or more diseases or conditions,such as for example, cancer.

Provided herein are methods of treating a cancer, in particular a solidtumor or a hematological cancer. The solid forms of Compound 1 describedherein provided herein can be used in the methods for treating orpreventing a cancer, in particular a solid tumor or a hematologicalcancer, as described herein. The methods comprise administering to asubject in need thereof an effective amount of a solid form of Compound1 described herein. Also provided herein are methods for treating andpreventing cancer metastasis, comprising administering to a subject inneed thereof an effective amount of a solid form of Compound 1 describedherein. The solid forms of Compound 1 described herein provided hereincan be used in the methods for treating and preventing cancermetastasis. Additionally, provided herein are methods of eradicatingcancer stem cells in a subject, comprising administering to a subject inneed thereof an effective amount of a solid form of Compound 1 describedherein. The solid forms of Compound 1 described herein provided hereincan be used in the methods of eradicating cancer stem cells in asubject. Also provided are methods of inducing differentiation in cancerstem cells in a subject, comprising administering to a subject in needthereof an effective amount of a solid form of Compound 1 describedherein. The solid forms of Compound 1 described herein provided hereincan be used in the methods of inducing differentiation in cancer stemcells in a subject. In another aspect, provided are methods of inducingcancer stem cell death in a subject, comprising administering to asubject in need thereof an effective amount of a solid form of Compound1 described herein. The solid forms of Compound 1 described hereinprovided herein can be used in the methods of inducing cancer stem celldeath in a subject.

Compounds useful in the methods disclosed herein include solid forms ofCompound 1 described herein, or a pharmaceutically acceptable salt,tautomer, stereoisomer, enantiomer, or isotopologue thereof.

The present embodiments can be understood more fully by reference to thedetailed description and examples, which are intended to exemplifynon-limiting embodiments.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a XRPD Stack Plot of Free Base Crystalline Forms A-I.

FIG. 2 depicts a XRPD Pattern of Free Base Form A.

FIG. 3 depicts a SEM Picture of Free Base Form A.

FIG. 4 depicts a TGA Thermogram of Free Base Form A.

FIG. 5 depicts a DSC Thermogram of Free Base Form A.

FIG. 6A depicts a DVS Isotherm Plot of Free Base Form A. FIG. 6B depictsthe values of the Isotherm Plot of FIG. 6A.

FIG. 7 depicts a ¹H NMR Spectrum of Free Base Form A.

FIG. 8 depicts a XRPD Pattern of Free Base Form B.

FIG. 9 depicts a TGA Thermogram of Free Base Form B.

FIG. 10 depicts a DSC Thermogram of Free Base Form B.

FIG. 11 depicts a XRPD Pattern of Free Base Form C.

FIG. 12 depicts a TGA Thermogram of Free Base Form C.

FIG. 13 depicts a DSC Thermogram of Free Base Form C.

FIG. 14 depicts a ¹H NMR Spectrum of Free Base Form C.

FIG. 15 depicts a XRPD Pattern of Free Base Form D.

FIG. 16 depicts a SEM Picture of Free Base Form D.

FIG. 17 depicts a TGA Thermogram of Free Base Form D.

FIG. 18 depicts a DSC Thermogram of Free Base Form D.

FIG. 19 depicts a ¹H NMR Spectrum of Free Base Form D

FIG. 20 depicts a XRPD Pattern of Free Base Form E.

FIG. 21 depicts a TGA Thermogram of Free Base Form E.

FIG. 22 depicts a DSC Thermogram of Free Base Form E.

FIG. 23 depicts a XRPD Pattern of Free Base Form F.

FIG. 24 depicts a SEM of Free Base Form F.

FIG. 25 depicts a TGA Thermogram of Free Base Form F.

FIG. 26 depicts a DSC Thermogram of Free Base Form F.

FIG. 27 depicts a ¹H NMR Spectrum of Free Base Form F.

FIG. 28 depicts a XRPD Pattern of Free Base Form G.

FIG. 29 depicts a SEM Picture of Free Base Form G.

FIG. 30 depicts a TGA Thermogram of Free Base Form G.

FIG. 31 depicts a DSC of Free Base Form G.

FIG. 32 depicts a ¹H NMR Spectrum of Free Base Form G.

FIG. 33 depicts a XRPD Pattern of Free Base Form H.

FIG. 34 depicts a TGA Thermogram of Free Base Form H.

FIG. 35 depicts a DSC Thermogram of Free Base Form H.

FIG. 36 depicts a XRPD Pattern of Free Base Form I.

FIG. 37 depicts a ¹H NMR Spectrum of Free Base Form I.

FIG. 38 depicts a TGA Thermogram of Free Base Form I.

FIG. 39 depicts a DSC Thermogram of Free Base Form I.

FIG. 40 depicts a XRPD Pattern of Amorphous Material.

FIG. 41 depicts a DSC Thermogram of Amorphous Material.

FIG. 42 depicts a ¹H NMR Spectrum of Amorphous Material.

FIG. 43A depicts a DVS Isotherm Plot of Amorphous Material. FIG. 43Bdepicts the values of the DVS Isotherm Plot of FIG. 43A.

FIG. 44 depicts a XRPD Stack Plot of Citrate Forms Y and Z.

FIG. 45 depicts a XRPD Pattern of Citrate Form Y.

FIG. 46 depicts a SEM Picture of Citrate Form Y.

FIG. 47 depicts a TGA Thermogram of Citrate Form Y.

FIG. 48 depicts a DSC Thermogram of Citrate Form Y.

FIG. 49A depicts a DVS Isotherm Plot Citrate Form Y. FIG. 49B depictsthe values of the Isotherm Plot of FIG. 49A.

FIG. 50 depicts a ¹H NMR Spectrum of Citrate Form Y.

FIG. 51A depicts a Comparison of XRPD Patterns of Citrate Form Y beforeCompression. FIG. 51B depicts a Comparison of XRPD Patterns of CitrateForm Y after Compression.

FIG. 52 depicts a XRPD Pattern of Citrate Form Z.

FIG. 53 depicts a SEM Picture of Citrate Form Z.

FIG. 54 depicts a TGA Thermogram of Citrate Form Z.

FIG. 55 depicts a DSC Thermogram of Citrate Form Z.

FIG. 56A depicts a DVS Isotherm Plot Citrate Form Z.

FIG. 56B depicts the values of the Isotherm Plot of FIG. 56A

FIG. 57 depicts a ¹H NMR Spectrum of Citrate Form Z.

FIG. 58 depicts a TGA Thermogram of a hydrate form of Citrate Form Z.

FIG. 59 depicts a TGA Thermogram of a non-stoichiometric form of CitrateForm Z.

FIG. 60 depicts a TGA Thermogram of a solvate form of Citrate Form Z.

FIG. 61 depicts ¹H NMR Spectrum of a solvate form of Citrate Form Z.

FIGS. 62A-62B depict a Comparison of XRPD Patterns of Citrate Form Z.FIG. 62A illustrates XRPD before compression. FIG. 62B illustrates XRPDafter compression.

FIG. 63 depicts a XRPD Pattern of HCl Salt starting material.

FIG. 64 depicts a TGA and DSC Thermogram of HCl Salt starting material.

FIG. 65 depicts a DVS Isotherm Plot of HCl Salt starting material.

FIG. 66 depicts a XRPD Pattern of HCl Salt Form 1.

FIG. 67 depicts a TGA and DSC Thermogram of HCl Salt Form 1.

FIG. 68 depicts a XRPD Pattern of HCl Salt Form 2.

FIG. 69 depicts a TGA and DSC Thermogram of HCl Salt Form 2.

FIG. 70 depicts a XRPD Pattern of HCl Salt Form 3.

FIG. 71 depicts a TGA and DSC Thermogram of HCl Salt Form 3.

FIG. 72 depicts a XRPD Pattern of HCl Salt Form 4.

FIG. 73 depicts a TGA and DSC Thermogram of HCl Salt Form 4.

FIG. 74 depicts a XRPD Pattern of HCl Salt Form 5.

FIG. 75 depicts a TGA and DSC Thermogram of HCl Salt Form 5.

FIG. 76 depicts a XRPD Pattern of HCl Salt Form 6.

FIG. 77 depicts a TGA and DSC Thermogram of HCl Salt Form 6.

FIG. 78 depicts a TGA Thermogram of HCl Salt Form 6.

FIG. 79 depicts a DSC Thermogram of HCl Salt Form 6.

FIG. 80 depicts a XRPD Pattern of HCl Salt Form 7.

FIG. 81 depicts a TGA and DSC Thermogram of HCl Salt Form 7.

FIG. 82 depicts a DSC Thermogram of HCl Salt Form 7.

FIG. 83 depicts a TGA Thermogram of HCl Salt Form 7.

FIG. 84 depicts a DVS Isotherm Plot of HCl Salt Form 7.

FIG. 85 depicts a XRPD Pattern of HCl Salt Form 8.

FIG. 86 depicts a TGA and DSC Thermogram of HCl Salt Form 8.

FIG. 87 depicts a TGA Thermogram of HCl Salt Form 8.

FIG. 88 depicts a DSC Thermogram of HCl Salt Form 8.

FIG. 89 depicts a XRPD Pattern of Compound 1.

FIG. 90 depicts a TGA Thermogram of Compound 1.

FIG. 91 depicts a DSC Thermogram of Compound 1.

FIG. 92 depicts a DVS Isotherm Plot of Compound 1.

FIG. 93 depicts a ¹H NMR of Compound 1.

FIG. 94 depicts a XRPD Pattern of Compound 1 HCl salt isolated fromsolubility study in SGF.

FIG. 95 depicts a TGA Thermogram of Compound 1 HCl salt isolated fromsolubility study in SGF.

FIG. 96 depicts a DSC Thermogram of Compound 1 HCl salt isolated fromsolubility study in SGF.

FIG. 97 depicts a DVS Isotherm Plot of Compound 1 HCl salt isolated fromsolubility study in SGF.

FIG. 98 depicts a ¹H NMR in D⁶-DMSO of Compound 1 HCl salt from SGFsolubility.

FIG. 99 depicts a ¹H NMR in D⁶-DMSO of Compound 1 sulfate salt from SVSSWell# H2.

FIG. 100 depicts a TGA Thermogram of sulfate salt SVSS Well# A2.

FIG. 101 depicts a DSC Thermogram of sulfate salt SVSS Well# A2.

FIG. 102 depicts a TGA Thermogram of sulfate salt SVSS Well# D2.

FIG. 103 depicts a DSC Thermogram of sulfate salt SVSS Well# D2.

FIG. 104 depicts a TGA Thermogram of sulfate salt SVSS Well# G2.

FIG. 105 depicts a DSC Thermogram of sulfate salt SVSS Well# A2.

FIG. 106 depicts a XRPD Pattern of mesylate salts from SVSS study inEtOAc.

FIG. 107 depicts a ¹H NMR in D⁶-DMSO of Compound 1 mesylate salt fromSVSS Well# B4.

FIG. 108 depicts a TGA Thermogram of mesylate salt SVSS Well# A4.

FIG. 109 depicts a DSC Thermogram of mesylate salt SVSS Well# A4.

FIG. 110 depicts a TGA Thermogram of mesylate salt SVSS Well# B4.

FIG. 111 depicts a DVS Isotherm Plot of mesylate salt SVSS Well# B4.

FIG. 112 depicts a TGA Thermogram of mesylate salt SVSS Well# E4.

FIG. 113 depicts a DSC Thermogram of mesylate salt SVSS Well# E4.

FIG. 114 depicts a TGA Thermogram of mesylate salt SVSS Well# G4.

FIG. 115 depicts a DSC Thermogram of mesylate salt SVSS Well# G4.

FIG. 116 depicts a TGA Thermogram of mesylate salt SVSS Well# H4.

FIG. 117 depicts a DSC Thermogram of mesylate salt SVSS Well# H4.

FIG. 118 depicts a XRPD Pattern of Compound 1 citrate salt from SVSS inethanol.

FIG. 119 depicts a XRPD Pattern of Compound 1 citrate salt from SVSS inIPA.

FIG. 120 depicts a XRPD Pattern of Compound 1 citrate salt from SVSS in3-methyl-2-butanol.

FIG. 121 depicts a XRPD Pattern of Compound 1 citrate salt from SVSS inacetonitrile.

FIG. 122 depicts a XRPD Pattern of Compound 1 citrate salt from SVSS inMTBE.

FIG. 123 depicts a XRPD Pattern of Compound 1 citrate salt from SVSS inacetone.

FIG. 124 depicts a XRPD Pattern of Compound 1 citrate salt from SVSS inwater.

FIG. 125 depicts a XRPD Pattern of Compound 1 citrate salt from SVSS inEtOAc.

FIG. 126 depicts XRPD comparison profiles of citrate salts from SVSS inethanol, IPA, MTBA, and acetone.

FIG. 127 depicts XRPD comparison profiles of citrate salts from SVSS in3-methyl-2-butanol and acetonitrile.

FIG. 128 depicts XRPD comparison profiles of citrate salts from SVSS inEtOAc and IPA.

FIG. 129 depicts a ¹H NMR in D⁶⁻DMSO of Compound 1 citrate salt fromSVSS Well# D9.

FIG. 130 depicts a TGA Thermogram of citrate salt from SVSS well# A9.

FIG. 131 depicts a DSC Thermogram of citrate salt from SVSS well# A9.

FIG. 132 depicts a TGA Thermogram of citrate salt from SVSS well# B9.

FIG. 133 depicts a TGA Thermogram of citrate salt from SVSS well# B9.

FIG. 134 depicts a TGA Thermogram of citrate salt from SVSS well# E9.

FIG. 135 depicts a DSC Thermogram of citrate salt from SVSS well# D9.

FIG. 136 depicts a TGA Thermogram of citrate salt from SVSS well# G9.

FIG. 137 depicts a DSC Thermogram of citrate salt from SVSS well# G9.

FIG. 138 depicts a TGA Thermogram of citrate salt from SVSS well# H9.

FIG. 139 depicts a DSC Thermogram of citrate salt from SVSS in EtOAcwell# H9.

FIG. 140 depicts a ¹H NMR in D⁶-DMSO of Compound 1 and phosphoric acidfrom SVSS Well# E7.

FIG. 141 depicts a ¹H NMR in D⁶-DMSO of Compound 1 and malic acid fromSVSS Well# G10.

FIG. 142 depicts a ¹H NMR in D⁶-DMSO of Compound 1 and glycolic acidfrom SVSS Well# G11.

FIG. 143 depicts a XRPD Pattern of HCl salt (top) compared with the onefrom solubility of free base in SGF (bottom).

FIG. 144 depicts a TGA Thermogram of HCl salt, monohydrate (1).

FIG. 145 depicts a DSC Thermogram of HCl salt, monohydrate (1).

FIG. 146 depicts a XRPD Pattern of HCl salt.

FIG. 147 depicts a TGA Thermogram of HCl salt.

FIG. 148 depicts a DSC Thermogram of HCl salt.

FIG. 149 depicts a XRPD Pattern of Compound 1 dried at 40° C. undervacuum.

FIG. 150 depicts a TGA Thermogram of Compound 1 dried at 40° C. undervacuum.

FIG. 151 depicts a DSC Thermogram of Compound 1 dried at 40° C. undervacuum.

FIG. 152 depicts a XRPD Pattern of Compound 1 HCl salt monohydrate afterheated to 140° C. on XRD-DSC stage.

FIG. 153 depicts a XRPD Pattern of Compound 1 sulfate.

FIG. 154 depicts a TGA Thermogram of Compound 1 sulfate.

FIG. 155 depicts a DSC Thermogram of Compound 1 sulfate.

FIG. 156 depicts a XRPD Pattern of Compound 1 mesylate.

FIG. 157 depicts a TGA Thermogram of Compound 1 mesylate.

FIG. 158 depicts a DSC Thermogram of Compound 1 mesylate.

FIG. 159 depicts a XRPD Pattern of Compound 1 mesylate.

FIG. 160 depicts a TGA Thermogram of Compound 1 mesylate.

FIG. 161 depicts a DSC Thermogram of Compound 1 mesylate.

FIG. 162 depicts a XRPD Pattern of Compound 1 mesylate after slurry inwater.

FIG. 163 depicts a DVS Isotherm Plot of Compound 1 mesylate salt.

FIG. 164 depicts a XRPD Pattern of Compound 1 mesylate after DVS study.

FIG. 165 depicts a XRPD Pattern of Compound 1 citrate in EtOAc-watersystem.

FIG. 166 depicts a TGA Thermogram of Compound 1 citrate in EtOAc-watersystem.

FIG. 167 depicts a DSC Thermogram of Compound 1 citrate in EtOAc-watersystem.

FIG. 168 depicts a XRPD Pattern of Compound 1 citrate in acetone.

FIG. 169 depicts a TGA Thermogram of Compound 1 citrate in acetone.

FIG. 170 depicts a DSC Thermogram of Compound 1 citrate in acetone.

FIG. 171 depicts a XRPD Pattern of Compound 1 citrate.

FIG. 172 depicts a TGA Thermogram of Compound 1 citrate.

FIG. 173 depicts a DSC Thermogram of Compound 1 citrate.

FIG. 174 depicts a XRPD Pattern of Compound 1 citrate.

FIG. 175 depicts a TGA Thermogram of Compound 1 citrate.

FIG. 176 depicts a DSC Thermogram of Compound 1 citrate.

FIG. 177 depicts a XRPD Pattern of Compound 1 citrate.

FIG. 178 depicts a TGA Thermogram of Compound 1 citrate.

FIG. 179 depicts a DSC Thermogram of Compound 1 citrate.

FIG. 180 depicts a XRPD Pattern of Compound 1 citrate.

FIG. 181 depicts a TGA Thermogram of Compound 1 citrate.

FIG. 182 depicts a DSC Thermogram of Compound 1 citrate.

FIG. 183 depicts a DVS Isotherm Plot of Compound 1 citrate salt.

FIG. 184 depicts a Dissolution of free base (FB), citrate and HCl saltin 0.01N HCl solution.

FIG. 185 depicts a Dissolution of Compound 1 free base (FB), citrate,and HCl salt in 0.001N HCl solution.

FIG. 186 depicts a Kinetic solubility of free base (FB), citrate, andHCl salt in FeSSIF.

FIG. 187 depicts a Kinetic solubility of free base (FB), citrate, andHCl salt in FaSSIF.

FIG. 188 illustrates Compound 1 Treatment Causes Sustained Inhibition ofthe ERK Substrate pRSK1 S380 in Colo 205 (mut BRAFV600E) Cells. Colo 205cells were treated with DMSO or 0.5 μM Compound 1 for indicated time.pRSK1 S380 was measured by MSD assay (Top). DUSP4 and DUSP6 weredetected by Western blotting (Bottom).

FIGS. 189A-189I illustrates Compound 1 potently inhibits MAP kinasesignaling and downstream target genes in Colo 205. Colon cancer cellline Colo 205 (BRAF V600E) cultures were treated with DMSO or increasingconcentrations of Compound 1 for 2, 8 or 24 h. FIG. 189A illustratesproteins extracted from treated cells and analyzed by Western blot usingantibodies against DUSP4, DUSP6, cyclin D1, c-Myc, YAP or β-actin. FIGS.189B-189C illustrate RNAs extracted using Cell-To-CT kit andquantitative PCR was performed with probes specific for DUSP4, DUSP6,SPRY2, c-Myc and cyclin D1. Specific probes for β-actin were used fornormalization. FIGS. 189D-189I illustrate Compound 1 Treatment modulatesMAPK-driven mRNA levels in Colo 205 (mut BRAFV600E) and HT-29 (mutBRAFV600E) Cells. Colo 205 or HT-29 cells were treated with DMSO or 0.3or 1 μM Compound 1 for 6 h. mRNA was extracted using MagMAX Total RNAIsolation kit and quantitative PCR was performed.

FIG. 190A illustrates Compound 1 effects on WNT/beta-catenin andHIPPO/YAP signaling pathway target genes in Colo 205. Colon cancer cellline Colo 205 (BRAF V600E) cultures were treated with DMSO or increasingconcentrations of Compound 1 for 2, 8 or 24 h. RNAs were extracted usingCell-To-CT kit and quantitative PCR was performed with probes specificfor Axin2, CTGF, and AREG. Specific probes for β-actin were used fornormalization. FIG. 190B-190E illustrate Compound 1 treatment regulatesYAP-driven mRNA levels in Colo 205 (mut BRAFV600E) and HT-29 (mutBRAFV600E) Cells. Colo 205 or HT-29 cells were treated with DMSO or 0.3or 1 μM Compound 1 for 6 h. RNAs were extracted using MagMAX Total RNAIsolation kit and quantitative PCR was performed.

FIGS. 191A-191B illustrate Compound 1 down-regulates PD-L1 level inmultiple cancer cell lines. FIG. 191A illustrates Western blotting oftotal PD-L1 in Hop66, Karpas-299, and LOX-IMVI. Cells were cultured inpresence or absence of Compound 1 for indicated time before expressionlevels of PD-L1, DUSP4 and α-tubulin or α-actin were measured by Westernblot. FIG. 191B illustrates surface staining of PD-L1 with theFluorescence-Activated Cell Sorter (FACS). Cells were treated with DMSOor Compound 1 at indicated concentrations for 48 h and cell surfaceexpression of PD-L1 was detected using the FACS analysis with anAPC-labeled antibody to PD-L1 (clone 29E.1A3; BioLegend, San Diego,Calif.). Geometric mean of PD-L1 positive cells was determined by FlowJo10 (Treestar, Ashland, Oreg.).

FIGS. 192A-192B illustrate Compound 1-treated KARPAS-299 cells increaseproduction of IL-2 (FIG. 192A) and IFNγ (FIG. 192B) by PBMC-derived Tcells stimulated with superantigen (SEB) in vitro. KARPAS-299 cells weretreated with DMSO (D) or Compound 1 at indicated concentrations for 48h. PBMC from healthy donors were treated with or without 20 ng/ml SEBfor 48 h. After wash with PBS, the PBMCs were incubated with the cancercells for 24 h and the supernatants were collected to measure IL-2 andIFNγ using MSD assays. FIG. 192C illustrates the effect of Compound 1treatment on levels of IL-8 were determined in PBMC culture media. PBMCswere isolated from whole blood and cultured in RPMI media plus 10% FBS.PBMCs were plated at 1×10⁶ per milliliter in 10 cm² dishes. The PBMCswere treated with 0.1% DMSO or 0.5 μM Compound 1. Treatments were takendown at designated time points. PBMCs were pelleted and used for Westernblot analysis and 1 mL of culture media was taken for IL-8 analysis. TheIL-8 analysis was performed with a Mesoscale V-Plex Human IL-8 kitaccording to the manufacturer's instructions. Compound 1 was shown toinhibit IL-8 levels at different time-points.

FIG. 193 illustrates antitumor activity of Compound 1 in the LOX-IMVIXenograft Model. Female SCID mice were inoculated with 1×10⁶ LOX-IMVItumor cells into the right flank. Mice were randomized into treatmentgroups (n=9/group) at the time of treatment initiation. Test articletreatment started on Day 13 when the tumors were approximately 240 mm.

FIG. 194 illustrates antitumor activity of Compound 1 in the LOX IMVIXenograft Model. Female severe-combined immunodeficient (SCID) mice wereinoculated with 1×10⁶ LOX-IMVI tumor cells into the right flank. Micewere randomized into treatment groups (n=10/group) at the time oftreatment initiation. Test article treatment started on Day 13 when thetumors were approximately 300 mm³. Percent inhibition is calculatedrelative to the vehicle control on the last study day and is inparentheses next to the respective tumor volume for the treatmentgroups. Dotted line is the tumor volume at the initiation of dosing.

FIG. 195 illustrates antitumor activity of Compound 1 in the Colo 205Xenograft Model. Female SCID mice were inoculated with 2×10⁶ Colo 205tumor cells into the right flank. Mice were randomized into treatmentgroups (n=10/group) at the time of treatment initiation. Test articletreatment started on Day 10 when the tumors were approximately 160 mm³.Percent inhibition is calculated relative to the vehicle control on thelast study day and is in parentheses next to the respective tumor volumefor the treatment groups. Dotted line is the tumor volume at theinitiation of dosing.

FIG. 196 illustrates antitumor activity of Compound 1 in the Colo 205Xenograft Model. Female SCID mice were inoculated with 2×10⁶ Colo 205tumor cells into the right flank. Mice were randomized into treatmentgroups (n=10/group) at the time of treatment initiation. Test articletreatment started on Day 10 when the tumors were approximately 130 or160 mm³. Percent inhibition is calculated relative to the vehiclecontrol on the last study day and is in parentheses next to therespective tumor volume for the treatment groups. Dotted line is thetumor volume at the initiation of dosing.

FIGS. 197A-197B illustrates antitumor activity of Compound 1 in thePDX146 Xenograft Model. Female NSG mice were inoculated with 25 μg ofPDX146 tumor in a cell slurry into the right flank. Mice were randomizedinto treatment groups (n=8-10/group) at the time of treatmentinitiation. Test article treatment started on Day 19 when the tumorswere approximately 100-110 mm³. FIG. 197A illustrates tumor volume as afunction of time. FIG. 10B illustrates individual tumor volume on thelast study day, day 40. Percent inhibition is calculated relative to thevehicle control on the last study day and is in parentheses next to therespective tumor volume for the treatment groups. Dotted line is thetumor volume at the initiation of dosing. Camp=camptosar.

FIG. 198 illustrates tumor Growth Delay with Continuous Compound 1Treatment in the PDX146 Xenograft Model. Female NSG mice were inoculatedwith 25 μg of PDX146 tumor in a cell slurry into the right flank. Micewere randomized into treatment groups (n=8-10/group) at the time oftreatment initiation. Test article treatment started on Day 16 when thetumors were approximately 100-110 mm³. Black dotted line is the tumorvolume at the initiation of dosing and the red dotted line is the tumorvolume on Day 43 when the vehicle control group was terminated.

FIGS. 199A-199D illustrates Single doses of Compound 1 inhibitbiomarkers in the MAPK, Wnt and Hippo signaling pathways in the PDX146Xenograft Model: Modulation of MAPK, Wnt and Hippo pathways in PDX146tumors treated with Compound 1. qRT-PCR assays were performed on RNAextracted from PDX146 tumors at the indicated time point post-dose. Dataare expressed as mean±SEM. P values are derived from a one-way ANOVAwith a Dunnet's post-hoc analysis.

FIGS. 200A-200D illustrate Compound 1 inhibits biomarkers in the MAPK,Wnt and Hippo signalling pathways from PDX146 tumors following a singledose administration: Modulation of MAPK, Wnt and Hippo pathways inPDX146 tumors treated with Compound 1. qRT-PCR assays were performed onRNA extracted from PDX146 tumors at the indicated time point post-dose.YAP data is generated from western blot analysis of tumors from the 5mg/kg treatment group and is expressed as a ratio of YAP to β-actinprotein expression. Data are expressed as mean±SEM. P values are derivedfrom a one-way ANOVA with a Dunnet's post-hoc analysis.

FIGS. 201A-201D illustrate phospho-RSK (pRSK) and phospho-ERK (pERK)protein levels, biomarkers of the MAPK signaling pathway, were modulatedby a single dose administration of Compound 1. Western blot (pRSK) orMesoscale (pERK) assays were performed on protein extracted from PDX146tumors at the indicated time point post-dose. Phospho-RSK data isexpressed as a % of the vehicle control. Phospho-ERK data is expressedas mean±SEM.

FIGS. 202A-202B illustrate antitumor activity of Compound 1 in theβ-catenin mutant SW48 colorectal xenograft model. Female SCID mice wereinoculated with 2×10⁶ SW48 tumor cells into the right flank. Mice wererandomized into treatment groups (n=10/group) at the time of treatmentinitiation. Test article treatment started on Day 10 when the tumorswere approximately 110 and 105 mm³ (FIG. 202A and FIG. 202B,respectively). Black dotted line is the tumor volume at the initiationof dosing. Graph on the left is a dose-response study (graph A). Graphon the right is a time to progression study where animals weremaintained on drug during the course of the study (graph B). Dotted lineis the tumor volume on Day 28 when the vehicle control group wasterminated.

FIG. 203 illustrates antitumor activity in the orthotopic Hep3B2.1-7hepatocellular carcinoma xenograft. Female SCID mice were orthotopicallyinoculated with 2×10⁶ Hep3B2.1-7 tumor cells per animal. Seven dayspost-inoculation animals were randomized into treatment groups based onbody weight and treatment commenced (Study day 0). Take rate assessmentof a satellite group confirmed the presence of tumor in the liver in100% of the animals. Compound 1 was dosed orally, QD for 21 days. On theday of study termination, tumors were removed and weighed. Individualtumor weights and the mean tumor weight±SEM of each group are plotted.Percent inhibition is calculated relative to the vehicle control and isabove the respective tumor weight for the treatment groups. P values arederived from a one-way ANOVA with a Dunnet's post-hoc analysis.***=p<0.001. Compound 1 showed a statistically significant reduction intumor weight compared to vehicle controls.

FIG. 204 illustrates antitumor activity of Compound 1 in the C-Metamplified hepatocellular carcinoma patient-derived xenograft model,LI0612. Female SCID mice were inoculated with hepatocellular carcinomaPDX model LI0612 tumor fragments (2-4 mm in diameter) into the rightflank. Mice were randomized into treatment groups (n=10/group) at thetime of treatment initiation. Test article treatment started on Day 18when the tumors were approximately 150 mm³. Tumor growth progressed inthe vehicle control and Compound 1 treatment groups over the dosingperiod. A change in the growth kinetics was noted with Compound 1administration resulting in significant tumor growth inhibition (TGI)with 30 mg/kg treatment (p=0.038, compared to the vehicle control).

FIG. 205 illustrates sensitivity of cell lines having β-cateninmutations to Compound 1 treatment and shows that cell lines with mutatedβ-catenin are generally more sensitive to Compound 1 treatment.

FIGS. 206A-206E illustrate cell line sensitivity and resistance totreatment with Compound 1. FIGS. 206A-206C show that cell linescontaining BRAF and CTNNB1 mutations are more sensitive to treatmentwith Compound 1 than cell lines with wild type BRAF and CTNNB1. FIG.206D and FIG. 206E show that cell lines with mutations in RB and thePI3K/PTEN pathway are associated with resistance to Compound 1 treatmentin vitro.

FIG. 207 illustrates Compound 1 modulates MAPK, β-catenin, and YAP inthe BRAF and CTNNB1 mutant cell line SW48.

FIGS. 208A-208B illustrate Compound 1 modulates target gene expressioncontrolled by MAPK, β-catenin, and YAP in the BRAF and CTNNB1 mutantcell line SW48.

FIG. 209 illustrates that Compound 1 inhibits Axin2 expression in humanbronchial epithelial cells. Gene expression was measured at 24 hours.

FIGS. 210A-210D illustrate that Compound 1 inhibits colony formation ofβ-catenin mutant cells at a level greater than MEK inhibitors(trametinib) and ERK inhibitors (GDC0994). FIG. 210A shows inhibition ofcolony formation of SW48 (colo) cells. FIG. 210B shows inhibition ofcolony formation of HCT-116 (colo) cells. FIG. 210C shows inhibition ofcolony formation of AGS (gastric) cells. FIG. 210D shows inhibition ofcolony formation of Hep3B (HCC) cells.

FIG. 211 illustrates that AGS cells resistant to the MEK inhibitortrametinib are sensitive to Compound 1 in a colony formation assay.

FIG. 212 illustrates TEAD reporter activity in 8×GTIIC-luciferase WI38VA13 cells treated with Compound 1 and trametinib for 72 hours.Luciferase activity was analyzed using the Bright Glo luciferase assay(Promega). Compound 1 inhibited TEAD reporter activity, with an averageIC₅₀ of >10 μM in the 24 hour assay and an average IC₅₀ of 1.85 μM inthe 72 hour assay (cumulative data of three experiments). Viability wasnot reproducibly affected by Compound 1 across the three assays.Trametinib did not inhibit TEAD reporter activity at 24 or 72 hours.

DETAILED DESCRIPTION Definitions

As used herein, and in the specification and the accompanying claims,the indefinite articles “a” and “an” and the definite article “the”include plural as well as single referents, unless the context clearlyindicates otherwise.

As used herein, and unless otherwise specified, the terms “about” and“approximately,” when used in connection with doses, amounts, or weightpercent of ingredients of a composition or a dosage form, mean a dose,amount, or weight percent that is recognized by one of ordinary skill inthe art to provide a pharmacological effect equivalent to that obtainedfrom the specified dose, amount, or weight percent. In certainembodiments, the terms “about” and “approximately,” when used in thiscontext, contemplate a dose, amount, or weight percent within 30%,within 20%, within 15%, within 10%, or within 5%, of the specified dose,amount, or weight percent.

As used herein, and unless otherwise specified, the terms “about” and“approximately,” when used in connection with a numeric value or rangeof values which is provided to characterize a particular solid form,e.g., a specific temperature or temperature range, such as, for example,that describes a melting, dehydration, desolvation, or glass transitiontemperature; a mass change, such as, for example, a mass change as afunction of temperature or humidity; a solvent or water content, interms of, for example, mass or a percentage; or a peak position, suchas, for example, in analysis by, for example, IR or Raman spectroscopyor XRPD; indicate that the value or range of values may deviate to anextent deemed reasonable to one of ordinary skill in the art while stilldescribing the solid form. Techniques for characterizing crystal formsand amorphous solids include, but are not limited to, thermalgravimetric analysis (TGA), differential scanning calorimetry (DSC),X-ray powder diffractometry (XRPD), single-crystal X-ray diffractometry,vibrational spectroscopy, e.g., infrared (IR) and Raman spectroscopy,solid-state and solution nuclear magnetic resonance (NMR) spectroscopy,optical microscopy, hot stage optical microscopy, scanning electronmicroscopy (SEM), electron crystallography and quantitative analysis,particle size analysis (PSA), surface area analysis, solubility studies,and dissolution studies. In certain embodiments, the terms “about” and“approximately,” when used in this context, indicate that the numericvalue or range of values may vary within 30%, 20%, 15%, 10%, 9%, 8%, 7%,6%, 5%, 4%, 3%, 2%, 1.5%, 1%, 0.5%, or 0.25% of the recited value orrange of values. For example, in some embodiments, the value of an XRPDpeak position may vary by up to ±0.1° 2θ (or ±0.2° 2θ) while stilldescribing the particular XRPD peak.

As used herein, and unless otherwise specified, a crystalline that is“pure,” i.e., substantially free of other crystalline or amorphoussolids, contains less than about 10% by weight of one or more othercrystalline or amorphous solids, less than about 5% by weight of one ormore other crystalline or amorphous solids, less than about 3% by weightof one or more other crystalline or amorphous solids, or less than about1% by weight of one or more other crystalline or amorphous solids.

As used herein, and unless otherwise specified, a solid form that is“substantially physically pure” is substantially free from other solidforms. In certain embodiments, a crystal form that is substantiallyphysically pure contains less than about 10%, 9%, 8%, 7%, 6%, 5%, 4%,3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.05%, or 0.01% of one or moreother solid forms on a weight basis. The detection of other solid formscan be accomplished by any method apparent to a person of ordinary skillin the art, including, but not limited to, diffraction analysis, thermalanalysis, elemental combustion analysis and/or spectroscopic analysis.

As used herein, and unless otherwise specified, a solid form that is“substantially chemically pure” is substantially free from otherchemical compounds (i.e., chemical impurities). In certain embodiments,a solid form that is substantially chemically pure contains less thanabout 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%,0.1%, 0.05%, or 0.01% of one or more other chemical compounds on aweight basis. The detection of other chemical compounds can beaccomplished by any method apparent to a person of ordinary skill in theart, including, but not limited to, methods of chemical analysis, suchas, e.g., mass spectrometry analysis, spectroscopic analysis, thermalanalysis, elemental combustion analysis and/or chromatographic analysis.

As used herein, and unless otherwise indicated, a chemical compound,solid form, or composition that is “substantially free” of anotherchemical compound, solid form, or composition means that the compound,solid form, or composition contains, in certain embodiments, less thanabout 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%0.1%, 0.05%, or 0.01% by weight of the other compound, solid form, orcomposition.

Unless otherwise specified, the terms “solvate” and “solvated,” as usedherein, refer to a solid form of a substance which contains solvent. Theterms “hydrate” and “hydrated” refer to a solvate wherein the solvent iswater. “Polymorphs of solvates” refer to the existence of more than onesolid form for a particular solvate composition. Similarly, “polymorphsof hydrates” refer to the existence of more than one solid form for aparticular hydrate composition. The term “desolvated solvate,” as usedherein, refers to a solid form of a substance which can be made byremoving the solvent from a solvate. The terms “solvate” and “solvated,”as used herein, can also refer to a solvate of a salt, cocrystal, ormolecular complex. The terms “hydrate” and “hydrated,” as used herein,can also refer to a hydrate of a salt, cocrystal, or molecular complex.

“Tautomers” refers to isomeric forms of a compound that are inequilibrium with each other. The concentrations of the isomeric formswill depend on the environment the compound is found in and may bedifferent depending upon, for example, whether the compound is a solidor is in an organic or aqueous solution. For example, in aqueoussolution, pyrazoles may exhibit the following isomeric forms, which arereferred to as tautomers of each other:

As readily understood by one skilled in the art, a wide variety offunctional groups and other structures may exhibit tautomerism and alltautomers of Compound 1 are within the scope of the present invention.

Unless otherwise specified, the term “composition” as used herein isintended to encompass a product comprising the specified ingredient(s)(and in the specified amount(s), if indicated), as well as any productwhich results, directly or indirectly, from combination of the specifiedingredient(s) in the specified amount(s). By “pharmaceuticallyacceptable,” it is meant a diluent, excipient, or carrier in aformulation must be compatible with the other ingredient(s) of theformulation and not deleterious to the recipient thereof.

The term “solid form” refers to a physical form which is notpredominantly in a liquid or a gaseous state. The terms “solid type” and“type” are used interchangeably herein with “solid form”. As used hereinand unless otherwise specified, the term “solid form,” when used hereinto refer to Compound 1, refers to a physical form comprising Compound 1which is not predominantly in a liquid or a gaseous state. A solid formmay be a crystalline form or a mixture thereof. In certain embodiments,a solid form may be a liquid crystal. In certain embodiments, the term“solid forms comprising Compound 1” includes crystal forms comprisingCompound 1. In certain embodiments, the solid form of Compound 1 is FormA, Form B, Form C, Form D, Form E, Form F, Form G, Form H, Form I, theamorphous solid, or a mixture thereof. In one embodiment, the solid formof Compound 1 is a citrate salt Form Y or citrate salt form Z. Incertain embodiments, the solid form of Compound 1 is HCl Salt Form 1,HCl Salt Form 2, HCl Salt Form 3, HCl Salt Form 4, HCl Salt Form 5, HClSalt Form 6, HCl Salt Form 7, HCl Salt Form 8, the amorphous solid, or amixture thereof.

As used herein and unless otherwise specified, the term “crystalline”when used to describe a compound, substance, modification, material,component or product, unless otherwise specified, means that thecompound, substance, modification, material, component or product issubstantially crystalline as determined by X-ray diffraction. See, e.g.,Remington: The Science and Practice of Pharmacy, 21st edition,Lippincott, Williams and Wilkins, Baltimore, Md. (2005); The UnitedStates Pharmacopeia, 23^(rd) ed., 1843-1844 (1995).

The term “crystal form” or “crystalline form” refers to a solid formthat is crystalline. In certain embodiments, a crystal form of asubstance may be substantially free of amorphous solids and/or othercrystal forms. In certain embodiments, a crystal form of a substance maycontain less than about 1%, less than about 2%, less than about 3%, lessthan about 4%, less than about 5%, less than about 6%, less than about7%, less than about 8%, less than about 9%, less than about 10%, lessthan about 15%, less than about 20%, less than about 25%, less thanabout 30%, less than about 35%, less than about 40%, less than about45%, or less than about 50% by weight of one or more amorphous solidsand/or other crystal forms. In certain embodiments, a crystal form of asubstance may be physically and/or chemically pure. In certainembodiments, a crystal form of a substance may be about 99%, about 98%,about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about91%, or about 90% physically and/or chemically pure.

Unless otherwise specified, the term “amorphous” or “amorphous solid”means that the substance, component, or product in question is notsubstantially crystalline as determined by X-ray diffraction. Inparticular, the term “amorphous solid” describes a disordered solidform, i.e., a solid form lacking long range crystalline order. Incertain embodiments, an amorphous solid of a substance may besubstantially free of other amorphous solids and/or crystal forms. Incertain embodiments, an amorphous solid of a substance may contain lessthan about 1%, less than about 2%, less than about 3%, less than about4%, less than about 5%, less than about 10%, less than about 15%, lessthan about 20%, less than about 25%, less than about 30%, less thanabout 35%, less than about 40%, less than about 45%, or less than about50% by weight of one or more other amorphous solids and/or crystal formson a weight basis. In certain embodiments, an amorphous solid of asubstance may be physically and/or chemically pure. In certainembodiments, an amorphous solid of a substance be about 99%, about 98%,about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about91%, or about 90% physically and/or chemically pure.

“Treating” as used herein, means an alleviation, in whole or in part, ofa disorder, disease or condition, or one or more of the symptomsassociated with a disorder, disease, or condition, or slowing or haltingof further progression or worsening of those symptoms, or alleviating oreradicating the cause(s) of the disorder, disease, or condition itself.In one embodiment, the disorder is a cancer, in particular, a solidtumor or hematological cancer. In some embodiments, “treating” means analleviation, in whole or in part, of a cancer, or symptoms associatedwith a cancer, in particular, a solid tumor or hematological cancer, ora slowing, or halting of further progression or worsening of thosesymptoms.

“Preventing” as used herein, means a method of delaying and/orprecluding the onset, recurrence or spread, in whole or in part, of acancer, in particular, a solid tumor or hematological cancer; barring asubject from acquiring a cancer, in particular, a solid tumor orhematological cancer; or reducing a subject's risk of acquiring acancer, in particular, a solid tumor or hematological cancer.

The term “effective amount” in connection with a solid form of Compound1 means an amount capable of treating or preventing a disorder, diseaseor condition, or symptoms thereof, disclosed herein. An effective amountrefers to an amount capable of treating or preventing a cancer, inparticular, a solid tumor or hematological cancer, or symptoms thereof,as disclosed herein. The effective amount of a solid form of Compound 1described herein, for example in a pharmaceutical composition, may be ata level that will exercise the desired effect; for example, about 0.005mg/kg of a subject's body weight to about 100 mg/kg of a patient's bodyweight in unit dosage for parenteral administration. As will be apparentto those skilled in the art, it is to be expected that the effectiveamount of a solid form of Compound 1 described herein may vary dependingon the severity of the indication being treated.

“Patient” or “subject” as used herein include an animal, including, butnot limited to, an animal such a cow, monkey, horse, sheep, pig,chicken, turkey, quail, cat, dog, mouse, rat, rabbit or guinea pig, inone embodiment a mammal, in another embodiment a human. In oneembodiment, a subject is a human having or at risk for having cancer, inparticular, a solid tumor or hematological cancer, or symptoms thereof.In one embodiment, a patient is a human having histologically orcytologically-confirmed solid tumor or hematological cancer, includingsubjects who have progressed on (or not been able to tolerate) standardanticancer therapy or for whom no standard anticancer therapy exists.

As used herein, and unless otherwise specified, the terms “cancer”refers to or describes the physiological condition in mammals that istypically characterized by unregulated cell growth. Examples of cancerinclude solid tumors and hematological cancer. In some embodiments, thecancer is a primary cancer, in others, the cancer is metastasized.

As used herein “solid tumors” includes, but is not limited to, bladdercancer (including, but not limited to, superficial bladder cancer),breast cancer (including, but not limited to, luminal B type, ER+, PR+and Her2+ breast cancer), central nervous system cancer (including, butnot limited to, glioblastoma multiforme (GBM), glioma, medulloblastoma,and astrocytoma), colorectal cancer, gastrointestinal cancer (including,but not limited to, stomach cancer, esophageal cancer, and rectumcancer), endocrine cancer (including, but not limited to, thyroidcancer, and adrenal gland cancer), eye cancer (including, but notlimited to, retinoblastoma), female genitourinary cancer (including, butnot limited to, cancer of the placenta, uterus, vulva, ovary, cervix),head and neck cancer (including, but not limited to, cancer of thepharynx, esophageal, and tongue), liver cancer, lung cancer (including,but not limited to, non-small cell lung cancer (NSCLC), small cell lungcancer (SCLC), mucoepidermoid, bronchogenic, squamous cell carcinoma(SQCC), and analplastic/NSCLC), skin cancer (including, but not limitedto, melanoma, and SQCC), soft tissue cancer (including but not limitedto, sarcoma, Ewing's sarcoma, and rhabdomyosarcoma), bone cancer(including, but not limited to, sarcoma, Ewing's sarcoma, andosteosarcoma), squamous cell cancer (including, but not limited to,lung, esophageal, cervical, and head and neck cancer), pancreas cancer,kidney cancer (including, but not limited to, renal Wilm's tumor andrenal cell carcinoma), and prostate cancer. In one embodiment, the solidtumor is not triple negative breast cancer (TNBC). In some embodiments,the solid tumor is breast cancer, colon cancer, lung cancer or bladdercancer. In one such embodiment, the solid tumor is superficial bladdercancer. In another, the solid tumor is lung squamous cell carcinoma. Inyet another embodiment, the solid tumor is luminal B type breast cancer.

As used herein “hematological cancer” includes, but is not limited to,leukemia (including, but not limited to, acute lymphocytic leukemia(ALL), chronic myeloid leukemia (CML), acute T-cell leukemia, B cellprecursor leukemia, acute promyelocytic leukemia (APML), plasma cellleukemia, myelomonoblastic/T-ALL, B myelomonocytic leukemia,erythroleukemia, and acute myeloid leukemia (AML)), lymphoma (includingbut not limited to Hodgkin's lymphoma, non-Hodgkin's lymphoma (NHL),Burkitt's lymphoma (BL), B cell lymphoma, lymphoblastic lymphoma,follicular lymphoma (FL), diffuse large B-cell lymphoma (DLBCL), largecell immunoblastic lymphoma), and multiple myeloma.

In the context of a cancer, inhibition may be assessed by inhibition ofdisease progression, inhibition of tumor growth, reduction of primarytumor, relief of tumor-related symptoms, inhibition of tumor secretedfactors (including tumor secreted hormones, such as those thatcontribute to carcinoid syndrome), delayed appearance of primary orsecondary tumors, slowed development of primary or secondary tumors,decreased occurrence of primary or secondary tumors, slowed or decreasedseverity of secondary effects of disease, arrested tumor growth andregression of tumors, increased Time To Progression (TTP), increasedProgression Free Survival (PFS), increased Overall Survival (OS), amongothers. OS as used herein means the time from treatment onset untildeath from any cause. TTP as used herein means the time from treatmentonset until tumor progression; TTP does not include deaths. As usedherein, PFS means the time from treatment onset until tumor progressionor death. In one embodiment, PFS rates will be computed using theKaplan-Meier estimates. In the extreme, complete inhibition, is referredto herein as prevention or chemoprevention. In this context, the term“prevention” includes either preventing the onset of clinically evidentcancer altogether or preventing the onset of a preclinically evidentstage of a cancer. Also intended to be encompassed by this definition isthe prevention of transformation into malignant cells or to arrest orreverse the progression of premalignant cells to malignant cells. Thisincludes prophylactic treatment of those at risk of developing a cancer.

In certain embodiments, the treatment of lymphoma may be assessed by theInternational Workshop Criteria (IWC) for non-Hodgkin lymphoma (NHL)(see Cheson B D, Pfistner B, Juweid, M E, et. al. Revised ResponseCriteria for Malignant Lymphoma. J. Clin. Oncol: 2007: (25) 579-586),using the response and endpoint definitions shown below:

Response Definition Nodal Masses Spleen, liver Bone Marrow CRDisappearance (a) FDG-avid or PET Not palpable, Infiltrate cleared on ofall evidence positive prior to nodules repeat biopsy; if of diseasetherapy; mass of any disappeared indeterminate by size permitted if PETmorphology, negative immunohistochemistry (b) Variably FDG-avid orshould be negative PET negative; regression to normal size on CT PRRegression of ≧50% decrease in SPD of ≧50% decrease in Irrelevant ifpositive measurable up to 6 largest dominant SPD of nodules prior totherapy; cell disease and no masses; no increase in (for single noduletype should be specified new sites size of other nodes in greatest (a)FDG-avid or PET transverse positive prior to diameter); no therapy; oneor more increase in size of PET positive at liver or spleen previouslyinvolved site (b) Variably FDG-avid or PET negative; regression on CT SDFailure to (a) FDG-avid or PET attain CR/PR or positive prior to PDtherapy; PET positive at prior sites of disease and no new sites on CTor PET (b) Variably FDG-avid or PET negative; no change in size ofprevious lesions on CT PD or Any new lesion Appearance of a new ≧50%increase New or recurrent relapsed or increase by lesion(s) ≧1.5 cm inany from nadir in the involvement disease ≧50% of axis, ≧50% increase inSPD of any previously SPD of more than one previous lesions involvedsites node, from nadir or ≧50% increase in longest diameter of apreviously identified node ≧1 cm in short axis Lesions PET positive ifFDG-avid lymphoma or PET positive prior to therapy

Abbreviations: CR, complete remission; FDG, [¹⁸F]fluorodeoxyglucose;PET, positron emission tomography; CT, computed tomography; PR, partialremission; SPD, sum of the product of the diameters; SD, stable disease;PD, progressive disease.

End point Patients Definition Measured from Primary Overall All Death asa result of any Entry onto study survival cause Progression- All Diseaseprogression or Entry onto study free survival death as a result of anycause Secondary Event-free All Failure of treatment or Entry onto studysurvival death as result of any cause Time to All Time to progression orEntry onto study progression death as a result of lymphoma Disease-freeIn CR Time to relapse or death Documentation survival as a result oflymphoma or of response acute toxicity of treatment Response In CR or PRTime to relapse or Documentation duration progression of responseLymphoma- All Time to death as a result Entry onto study specific oflymphoma survival Time to All Time to new treatment End of primary nexttreatment treatment Abbreviations: CR: complete remission; PR: partialremission.

In one embodiment, the end point for lymphoma is evidence of clinicalbenefit. Clinical benefit may reflect improvement in quality of life, orreduction in patient symptoms, transfusion requirements, frequentinfections, or other parameters. Time to reappearance or progression oflymphoma-related symptoms can also be used in this end point.

In certain embodiments, the treatment of CLL may be assessed by theInternational Workshop Guidelines for CLL (see Hallek M, Cheson B D,Catovsky D, et al. Guidelines for the diagnosis and treatment of chroniclymphocytic leukemia: a report from the International Workshop onChronic Lymphocytic Leukemia updating the National CancerInstitute-Working Group 1996 guidelines. Blood, 2008; (111) 12:5446-5456) using the response and endpoint definitions shown therein andin particular:

Parameter CR PR PD Group A Lymphadenopathy^(†) None >1.5 cm Decrease≧50% Increase ≧50% Hepatomegaly None Decrease ≧50% Increase ≧50%Splenomegaly None Decrease ≧50% Increase ≧50% Blood lymphocytes <4000/μLDecrease ≧50% Increase ≧50% from baseline over baseline Marrow‡Normocellular, 50% reduction <30% in marrow lymphocytes, infiltrate, orno B-lymphoid B-lymphoid nodules. nodules Hypocellular marrow definesCRi (5.1.6). Group B Platelet count >100 000/μL >100 000/μL or Decreaseof increase ≧50% ≧50% from over baseline baseline secondary to CLLHemoglobin >11.0 g/dL >11 g/dL or Decrease of increase ≧50% >2 g/dL fromover baseline baseline secondary to CLLNeutrophils^(‡) >1500/μL >1500/μL or >50% improvement over baseline

Group A criteria define the tumor load; Group B criteria define thefunction of the hematopoietic system (or marrow). CR (completeremission): all of the criteria have to be met, and patients have tolack disease-related constitutional symptoms; PR (partial remission): atleast two of the criteria of group A plus one of the criteria of group Bhave to be met; SD is absence of progressive disease (PD) and failure toachieve at least a PR; PD: at least one of the above criteria of group Aor group B has to be met. Sum of the products of multiple lymph nodes(as evaluated by CT scans in clinical trials, or by physical examinationin general practice). These parameters are irrelevant for some responsecategories.

In certain embodiments, the treatment of multiple myeloma may beassessed by the International Uniform Response Criteria for MultipleMyeloma (IURC) (see Durie B G M, Harousseau J-L, Miguel J S, et al.International uniform response criteria for multiple myeloma. Leukemia,2006; (10) 10: 1-7), using the response and endpoint definitions shownbelow:

Response Subcategory Response Criteria^(a) sCR CR as defined below plusNormal FLC ratio and Absence of clonal cells in bone marrow^(b) byimmunohistochemistry or immunofluorescence^(c) CR Negativeimmunofixation on the serum and urine and Disappearance of any softtissue plasmacytomas and <5% plasma cells in bone marrow^(b) VGPR Serumand urine M-protein detectable by immunofixation but not onelectrophoresis or 90% or greater reduction in serum M-protein plusurine M-protein level <100 mg per 24 h PR ≧50% reduction of serumM-protein and reduction in 24-h urinary M-protein by ≧90% or to <200 mgper 24 h If the serum and urine M-protein are unmeasurable, ^(d) a ≧50%decrease in the difference between involved and uninvolved FLC levels isrequired in place of the M- protein criteria If serum and urineM-protein are unmeasurable, and serum free light assay is alsounmeasurable, ≧50% reduction in plasma cells is required in place ofM-protein, provided baseline bone marrow plasma cell percentage was ≧30%In addition to the above listed criteria, if present at baseline, a ≧50%reduction in the size of soft tissue plasmacytomas is also required SD(not Not meeting criteria for CR, VGPR, PR or recommended forprogressive disease use as an indicator of response; stability ofdisease is best described by providing the time to progressionestimates)

Abbreviations: CR, complete response; FLC, free light chain; PR, partialresponse; SD, stable disease; sCR, stringent complete response; VGPR,very good partial response; ^(a)All response categories require twoconsecutive assessments made at any time before the institution of anynew therapy; all categories also require no known evidence ofprogressive or new bone lesions if radiographic studies were performed.Radiographic studies are not required to satisfy these responserequirements; ^(b)Confirmation with repeat bone marrow biopsy notneeded; ^(c)Presence/absence of clonal cells is based upon the κ/λratio. An abnormal κ/λ ratio by immunohistochemistry and/orimmunofluorescence requires a minimum of 100 plasma cells for analysis.An abnormal ratio reflecting presence of an abnormal clone is κ/λof >4:1 or <1:2. ^(d)Measurable disease defined by at least one of thefollowing measurements: Bone marrow plasma cells ≧30%; Serum M-protein≧1 g/dl (≧10 gm/l)[10 g/l]; Urine M-protein ≧200 mg/24 h; Serum FLCassay: Involved FLC level ≧10 mg/dl (≧100 mg/l); provided serum FLCratio is abnormal.

In certain embodiments, the treatment of a cancer may be assessed byResponse Evaluation Criteria in Solid Tumors (RECIST 1.1) (see ThereasseP., et al. New Guidelines to Evaluate the Response to Treatment in SolidTumors. J. of the National Cancer Institute; 2000; (92) 205-216 andEisenhauer E. A., Therasse P., Bogaerts J., et al. New responseevaluation criteria in solid tumors: Revised RECIST guideline (version1.1). European J. Cancer; 2009; (45) 228-247). Overall responses for allpossible combinations of tumor responses in target and non-targetlesions with our without the appearance of new lesions are as follows:

Target lesions Non-target lesions New lesions Overall response CR CR NoCR CR Incomplete No PR response/SD PR Non-PD No PR SD Non-PD No SD PDAny Yes or no PD Any PD Yes or no PD Any Any Yes PD CR = completeresponse; PR = partial response; SD = stable disease; and PD =progressive disease.

With respect to the evaluation of target lesions, complete response (CR)is the disappearance of all target lesions, partial response (PR) is atleast a 30% decrease in the sum of the longest diameter of targetlesions, taking as reference the baseline sum longest diameter,progressive disease (PD) is at least a 20% increase in the sum of thelongest diameter of target lesions, taking as reference the smallest sumlongest diameter recorded since the treatment started or the appearanceof one or more new lesions and stable disease (SD) is neither sufficientshrinkage to qualify for partial response nor sufficient increase toqualify for progressive disease, taking as reference the smallest sumlongest diameter since the treatment started.

With respect to the evaluation of non-target lesions, complete response(CR) is the disappearance of all non-target lesions and normalization oftumor marker level; incomplete response/stable disease (SD) is thepersistence of one or more non-target lesion(s) and/or the maintenanceof tumor marker level above the normal limits, and progressive disease(PD) is the appearance of one or more new lesions and/or unequivocalprogression of existing non-target lesions.

The procedures, conventions, and definitions described below provideguidance for implementing the recommendations from the ResponseAssessment for Neuro-Oncology (RANO) Working Group regarding responsecriteria for high-grade gliomas (Wen P., Macdonald, D R., Reardon, D A.,et al. Updated response assessment criteria for high-grade gliomas:Response assessment in neuro-oncology working group. J Clin Oncol 2010;28: 1963-1972). Primary modifications to the RANO criteria for Criteriafor Time Point Responses (TPR) can include the addition of operationalconventions for defining changes in glucocorticoid dose, and the removalof subjects' clinical deterioration component to focus on objectiveradiologic assessments. The baseline MM scan is defined as theassessment performed at the end of the post-surgery rest period, priorto initiating or re-initiating compound treatment. The baseline MRI isused as the reference for assessing complete response (CR) and partialresponse (PR). Whereas, the smallest SPD (sum of the products ofperpendicular diameters) obtained either at baseline or at subsequentassessments will be designated the nadir assessment and utilized as thereference for determining progression. For the 5 days preceding anyprotocol-defined MRI scan, subjects receive either no glucocorticoids orare on a stable dose of glucocorticoids. A stable dose is defined as thesame daily dose for the 5 consecutive days preceding the MRI scan. Ifthe prescribed glucocorticoid dose is changed in the 5 days before thebaseline scan, a new baseline scan is required with glucocorticoid usemeeting the criteria described above. The following definitions will beused.

Measurable Lesions: Measurable lesions are contrast-enhancing lesionsthat can be measured bi-dimensionally. A measurement is made of themaximal enhancing tumor diameter (also known as the longest diameter,LD). The greatest perpendicular diameter is measured on the same image.The cross hairs of bi-dimensional measurements should cross and theproduct of these diameters will be calculated.

Minimal Diameter: T1-weighted image in which the sections are 5 mm with1 mm skip. The minimal LD of a measurable lesion is set as 5 mm by 5 mm.Larger diameters may be required for inclusion and/or designation astarget lesions. After baseline, target lesions that become smaller thanthe minimum requirement for measurement or become no longer amenable tobi-dimensional measurement will be recorded at the default value of 5 mmfor each diameter below 5 mm. Lesions that disappear will be recorded as0 mm by 0 mm.

Multicentric Lesions: Lesions that are considered multicentric (asopposed to continuous) are lesions where there is normal interveningbrain tissue between the two (or more) lesions. For multicentric lesionsthat are discrete foci of enhancement, the approach is to separatelymeasure each enhancing lesion that meets the inclusion criteria. Ifthere is no normal brain tissue between two (or more) lesions, they willbe considered the same lesion.

Nonmeasurable Lesions: All lesions that do not meet the criteria formeasurable disease as defined above will be considered non-measurablelesions, as well as all non-enhancing and other truly nonmeasurablelesions. Nonmeasurable lesions include foci of enhancement that are lessthan the specified smallest diameter (i.e., less than 5 mm by 5 mm),non-enhancing lesions (e.g., as seen on T1-weighted post-contrast,T2-weighted, or fluid-attenuated inversion recovery (FLAIR) images),hemorrhagic or predominantly cystic or necrotic lesions, andleptomeningeal tumor. Hemorrhagic lesions often have intrinsicT1-weighted hyperintensity that could be misinterpreted as enhancingtumor, and for this reason, the pre-contrast T1-weighted image may beexamined to exclude baseline or interval sub-acute hemorrhage.

At baseline, lesions will be classified as follows: Target lesions: Upto 5 measurable lesions can be selected as target lesions with eachmeasuring at least 10 mm by 5 mm, representative of the subject'sdisease; Non-target lesions: All other lesions, including allnonmeasurable lesions (including mass effects and T2/FLAIR findings) andany measurable lesion not selected as a target lesion. At baseline,target lesions are to be measured as described in the definition formeasurable lesions and the SPD of all target lesions is to bedetermined. The presence of all other lesions is to be documented. Atall post-treatment evaluations, the baseline classification of lesionsas target and non-target lesions will be maintained and lesions will bedocumented and described in a consistent fashion over time (e.g.,recorded in the same order on source documents and eCRFs). Allmeasurable and nonmeasurable lesions must be assessed using the sametechnique as at baseline (e.g., subjects should be imaged on the same MMscanner or at least with the same magnet strength) for the duration ofthe study to reduce difficulties in interpreting changes. At eachevaluation, target lesions will be measured and the SPD calculated.Non-target lesions will be assessed qualitatively and new lesions, ifany, will be documented separately. At each evaluation, a time pointresponse will be determined for target lesions, non-target lesions, andnew lesion. Tumor progression can be established even if only a subsetof lesions is assessed. However, unless progression is observed,objective status (stable disease, PR or CR) can only be determined whenall lesions are assessed.

Confirmation assessments for overall time point responses of CR and PRwill be performed at the next scheduled assessment, but confirmation maynot occur if scans have an interval of <28 days. Best response,incorporating confirmation requirements, will be derived from the seriesof time points.

Compound 1

The solid forms, formulations and methods of use provided herein relateto solid forms (e.g., polymorphs) of Compound 1:

having the name(1s,4s)-4-(8-((2,4,6-trichlorophenyl)amino)-2-(((3S,4R)-3-fluorotetrahydro-2H-pyran-4-yl)amino)-9H-purin-9-yl)-1-methylcyclohexane-1-carboxamide,alternatively namedcis-4-[2-{[(3S,4R)-3-fluorooxan-4-yl]amino}-8-(2,4,6-trichloroanilino)-9H-purin-9-yl]-1-methylcyclohexane-1-carboxamide,including tautomers thereof.

Solid Forms of Compound 1

In certain embodiments, provided herein are solid forms of Compound 1.In certain embodiments, the solid form is crystalline. In certainembodiments, the solid form is a single-component solid form. In certainembodiments, the solid form is a hydrate. In certain embodiments, thesolid form is an anhydrate. In certain embodiments, the solid form is anHCl salt of Compound 1. In certain embodiments, the solid form is acitrate salt of Compound 1. In certain embodiments, the solid form is amesylate salt. In certain embodiments, the solid form is a sulfate salt.In certain embodiments, the solid form is a solvate.

While not intending to be bound by any particular theory, certain solidforms are characterized by physical properties, e.g., stability,solubility and dissolution rate, appropriate for pharmaceutical andtherapeutic dosage forms. Moreover, while not wishing to be bound by anyparticular theory, certain solid forms are characterized by physicalproperties (e.g., density, compressibility, hardness, morphology,cleavage, stickiness, solubility, water uptake, electrical properties,thermal behavior, solid-state reactivity, physical stability, andchemical stability) affecting particular processes (e.g., yield,filtration, washing, drying, milling, mixing, tableting, flowability,dissolution, formulation, and lyophilization) which make certain solidforms suitable for the manufacture of a solid dosage form. Suchproperties can be determined using particular analytical chemicaltechniques, including solid-state analytical techniques (e.g., X-raydiffraction, microscopy, spectroscopy and thermal analysis), asdescribed herein and known in the art.

The solid forms provided herein (e.g., Form A, Form B, Form C, Form D,Form E, Form F, Form G, Form H, Form I, and the amorphous solid ofCompound 1, and HCl salt Form 1, HCl Salt Form 2, HCl Salt Form 3, HClSalt Form 4, HCl Salt Form 5, HCl Salt Form 6, HCl Salt Form 7, HCl SaltForm 8, and the HCl salt amorphous solid of Compound 1, and citrate saltForm Y, Form Z and the citrate salt amorphous solid of Compound 1) maybe characterized using a number of methods known to a person skilled inthe art, including, but not limited to, single crystal X-raydiffraction, X-ray powder diffraction (XRPD), microscopy (e.g., scanningelectron microscopy (SEM)), thermal analysis (e.g., differentialscanning calorimetry (DSC), dynamic vapor sorption (DVS), thermalgravimetric analysis (TGA), and hot-stage microscopy), spectroscopy(e.g., infrared, Raman, and solid-state nuclear magnetic resonance),ultra-high performance liquid chromatography (UHPLC), and proton nuclearmagnetic resonance NMR) spectrum. The particle size and sizedistribution of the solid form provided herein may be determined byconventional methods, such as laser light scattering technique.

The purity of the solid forms provided herein may be determined bystandard analytical methods, such as thin layer chromatography (TLC),gel electrophoresis, gas chromatography, ultra-high performance liquidchromatography (UHPLC), and mass spectrometry (MS).

It should be understood that the numerical values of the peaks of anX-ray powder diffraction pattern may vary slightly from one machine toanother or from one sample to another, and so the values quoted are notto be construed as absolute, but with an allowable variability, such as±0.2° 2θ or ±0.1° 2θ (see United State Pharmacopoeia, page 2228 (2003)).

In certain embodiments, provided herein are methods for making a solidform of Compound 1, comprising 1) obtaining a slurry of Compound 1 in asolvent; 2) stirring the slurry for a period of time (e.g., about 24 h)at a certain temperature (e.g., about 25° C. or about 50° C.); and 3)collecting solids from the slurry by filtration and optionally drying,where the solids can be Form A. In certain embodiments, provided hereinare methods for making a solid form of Compound 1, comprising 1)obtaining a slurry of Compound 1 in a solvent; 2) stirring the slurryfor about 24 h at about 25° C. or about 50° C.; and 3) collecting solidsfrom the slurry by filtration, for example through a 0.45 μm PTFEsyringe filter and optionally air drying, where the solids can be FormA. In certain embodiments, the methods for making a solid form ofCompound 1 are equilibration experiments, such as slurry experiments.

In certain embodiments, provided herein are methods for making a solidform of Compound 1, comprising 1) dissolving Compound 1 in a solvent toyield a solution; 2) filtering the solution if the compound does notdissolve completely; and 3) evaporating the solution under certain airpressure (e.g., about 1 atm) at a certain temperature (e.g., about 25°C. or about 50° C.) to yield a solid that is optionally Form A. Incertain embodiments, provided herein are methods for making a solid formof Form A, comprising 1) dissolving Compound 1 in a solvent to yield asolution; 2) filtering the solution (for example, through a 0.45 μm PTFEsyringe filter) if Form A does not dissolve completely; and 3)evaporating the solution under about 1 atm air pressure at about 25° C.or about 50° C. under nitrogen to yield a solid. In certain embodiments,the methods for making a solid form of Compound 1 are evaporationexperiments.

In certain embodiments, provided herein are methods for making a solidform of Compound 1, comprising 1) obtaining a saturated solution of FormA in a solvent at a first temperature (e.g., about 60° C.); 2) stirringthe solution at the first temperature for a period of time (e.g., 10minutes); 3) filtering the solution; 4) cooling the solution slowly to asecond temperature (e.g., about −5° C. to about 15° C.); and 5)isolating solids from the solution and optionally drying. In certainembodiments, provided herein are methods for making a solid form ofCompound 1, comprising 1) obtaining a saturated solution of Form A in asolvent at about 60° C.; 2) stirring the solution at about 60° C. for 10minutes; 3) filtering the solution (for example through a 0.45 μm PTFEsyringe filter); 4) cooling the solution slowly to about 5° C.; and 5)isolating solids from the solution and optionally air-drying. In certainembodiments, the methods for making a solid form of Compound 1 arecooling recrystallization experiments.

In certain embodiments, provided herein are methods for making a solidform of Compound 1, comprising 1) obtaining a saturated solution of FormA in a solvent at a first temperature (e.g., about 60° C.); 2) adding ananti-solvent into the saturated solution at the first temperature; 3)cooling down to a second temperature (e.g., about −5° C. to about 15°C.); and 4) collecting a solid if there is precipitation, andevaporating the solvent to collect a solid if there is no precipitation;and 5) optionally drying. In certain embodiments, provided herein aremethods for making a solid form of Compound 1, comprising 1) obtaining asaturated solution of Form A in a solvent at about 60° C.; 2) adding ananti-solvent into the saturated solution at about 60° C.; 3) coolingdown to about 5° C.; and 4) collecting a solid if there isprecipitation, and evaporating the solvent to collect a solid if thereis no precipitation; and 5) optionally air drying. In certainembodiments, the ratio by volume of solvent and anti-solvent is about1:9. In certain embodiments, the methods for making a solid form ofCompound 1 are anti-solvent recrystallization experiments.

In certain embodiments, the solvent is acetone, DCM, EtOAc, EtOH,EtOH/H₂O (about 1:1), H₂O, heptane, IPA, ACN, ACN/H₂O (about 1:1), MEK,MeOH, MTBE, n-BuOH, THF, THF/H₂O (about 1:1), toluene or sulfolane.

In certain embodiments, the anti-solvent is ACN, heptane, MTBE, orwater.

Form A

In certain embodiments, provided herein is Form A.

In one embodiment, Form A is a solid form of Compound 1. In oneembodiment, Form A is a monohydrate. In one embodiment, Form A is anon-stoichiometric channel hydrate solid form of Compound 1. In oneembodiment, Form A is a free base form of Compound 1. In anotherembodiment, Form A is crystalline.

In certain embodiments, Form A provided herein is obtained byequilibration experiments, evaporation experiments and anti-solventrecrystallization experiments (see Table 6, Table 7, and Table 9). Incertain embodiments, Form A is obtained from certain solvent systemsincluding heptane/water, heptanes, water, toluene, MeCN, MeCN/water,EtOH, EtOH/H₂O (about 1:1), THF/water (about 1:1), and IPA.

In one embodiment, a method of preparing Form A comprises the steps ofcontacting Compound 1 (e.g., a crystalline form of Compound 1 such asForm B, Form C, or Form H) with ambient conditions comprising greaterthan about 10%-20% relative humidity (RH).

In one embodiment, a method of preparing Form A comprises the steps ofcooling Compound 1 in a solvent to a temperature less than about 50° C.and collecting solids.

In certain embodiments, a solid form provided herein, e.g., Form A, isthe free base of Compound 1, and is substantially crystalline, asindicated by, e.g., X-ray powder diffraction measurements. In oneembodiment, is an X-ray powder diffraction pattern (XRPD) substantiallyas shown in FIG. 2 (e.g. Form A). In one embodiment, a solid formprovided herein, e.g., Form A, has one or more characteristic X-raypowder diffraction peaks at approximately 3.2, 7.3, 8.5, 10.7, 11.1,12.7, 13.0, 13.4, 13.8, 14.5, 14.7, 15.9, 16.9, 17.1, 17.3, 17.7, 18.2,18.7, 20.3, 20.7, 21.0, 21.3, 22.1, 22.7, 22.9, 23.2, 23.6, 24.0, 24.8,25.5, 26.1, 26.4, 26.8, 27.9, 28.1, 28.8, 29.4, 29.8, 31.4, 31.8, 32.6,33.1, 33.6, 33.9, 34.2, 34.7, 36.1, 36.5, 37.2, 37.7, 38.9, or 39.5,° 2θ(±0.2° 2θ) or (±0.1° 2θ) as depicted in FIG. 2. In a specificembodiment, a solid form provided herein has one, two, three, four,five, six, seven, eight, nine, ten, eleven, or twelve characteristicX-ray powder diffraction peaks at approximately 7.3, 8.5, 10.8, 14.5,14.7, 15.9, 16.9, 17.1, 18.2, 21.0, 21.3, or 28.8° 2θ (±0.2° 2θ). Inembodiments, the solid form is Form A. In another embodiment, a solidform provided herein has one, two, three or four characteristic X-raypowder diffraction peaks at approximately 7.3, 8.5, 18.2, or 21.3° 2θ(±0.2° 2θ). In another embodiment, Form A has one, two, three, four,five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen,fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one,twenty-two, twenty-three, twenty-four, twenty-five, twenty-six,twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two,thirty-three, thirty-four, thirty-five, thirty-six, thirty-seven,thirty-eight, thirty-nine, forty, forty-one, forty-two, forty-three,forty-four, forty-five, forty-six, forty-seven, forty-eight, forty-nine,fifty, fifty-one, or fifty-two characteristic X-ray powder diffractionpeaks as set forth in Table 12.

In one embodiment, a solid form provided herein, e.g. Form A, has a SEMimage substantially as shown in FIG. 3.

In one embodiment, provided herein is a solid form, e.g. Form A, havinga TGA thermograph corresponding substantially to the representative TGAthermogram as depicted in FIG. 4. In certain embodiments, thecrystalline form exhibits a TGA thermogram comprising a total mass lossof approximately 2.8% of the total mass of the sample betweenapproximately 50° C. and approximately 175° C. when heated fromapproximately 50° C. to approximately 220° C. Thus, in certainembodiments, the crystalline form loses about 2.8% its total mass whenheated from about ambient temperature to about 220° C.

In one embodiment, provided herein is a solid form, e.g. Form A, havinga DSC thermogram substantially as depicted in FIG. 5 comprising anendothermic event with an onset temperature of about 94° C. and a peakmaximum temperature of about 117° C. In one embodiment, provided hereinis a solid form, e.g. Form A, having a DSC thermogram substantially asdepicted in FIG. 5 comprising an endothermic event with an onsettemperature of about 174° C. and a peak maximum temperature of about182° C. when heated from approximately 25° C. to approximately 220° C.

In one embodiment, provided herein is a solid form, e.g. Form A, havinga DVS isotherm plot substantially as depicted in FIG. 6A.

In one embodiment, provided herein is Form A having a ¹H NMR spectrumsubstantially as depicted in FIG. 7.

In still another embodiment, Form A is substantially pure. In certainembodiments, the substantially pure Form A is substantially free ofother solid forms, e.g., amorphous solid. In certain embodiments, thesubstantially pure Form A is substantially free of Form B, Form C, orForm H. In certain embodiments, the purity of the substantially pureForm A is no less than about 95%, no less than about 96%, no less thanabout 97%, no less than about 98%, no less than about 98.5%, no lessthan about 99%, no less than about 99.5%, or no less than about 99.8%.

Form B

In certain embodiments, provided herein is Form B.

In one embodiment, Form B is a solid form of Compound 1. In anotherembodiment, Form B is crystalline. In one embodiment, Form B is ananhydrate form of Compound 1.

In certain embodiments, Form B provided herein is obtained byequilibration experiments, evaporation experiments and anti-solventrecrystallization experiments (see Table 6, Table 7, and Table 9). Incertain embodiments, Form B is obtained from certain solvent systemsincluding heptane/water, heptanes, water, toluene, MeCN, MeCN/water,EtOH, EtOH/H₂O (about 1:1), THF/water (about 1:1), and IPA. In certainembodiments, Form B is obtained by drying or reducing the RH subjectedto Form A to less than about 10%.

In certain embodiments, a solid form provided herein, e.g., Form B, issubstantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, Form B has an X-ray powderdiffraction pattern substantially as shown in FIG. 8. In one embodiment,a solid form provided herein, e.g. Form B, has one or morecharacteristic X-ray powder diffraction peaks at approximately 6.9, 8.7,10.5, 11.6, 12.0, 13.6, 13.8, 14.1, 14.2, 16.3, 16.9, 17.5, 18.0, 18.4,19.1, 19.5, 20.0, 20.8, 21.1, 22.1, 22.7, 23.3, 25.2, 26.0, 26.7, 27.4,28.4, 28.8, 29.2, 30.1, 31.0, 31.5, or 31.8° 2θ (±0.2° 2θ) or (±0.1° 2θ)as depicted in FIG. 8. In a specific embodiment, a solid form providedherein has one, two, three, four, five, six, seven, eight, nine, or tencharacteristic X-ray powder diffraction peaks at approximately 8.7,11.6, 12.0, 13.8, 14.1, 17.5, 18.0, 19.5, 20.0, or 20.8° 2θ (±0.2° 2θ).In a specific embodiment, a solid form provided herein has one, two,three, or four characteristic X-ray powder diffraction peaks atapproximately 13.8, 19.5, 20.0, or 20.8° 2θ (±0.2° 2θ). In oneembodiment, the solid form is Form B. In another embodiment, Form B hasone, two, three, four, five, six, seven, eight, nine, ten, eleven,twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen,nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four,twenty-five, twenty-six, twenty-seven, twenty-eight, twenty-nine,thirty, thirty-one, thirty-two, or thirty-three characteristic X-raypowder diffraction peaks as set forth in Table 13.

In one embodiment, provided herein is a crystalline form of Compound 1,e.g. Form B, having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 9. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 0.1% of the total mass of the samplebetween approximately 30° C. and approximately 155° C. when heated fromapproximately 25° C. to approximately 220° C. Thus, in certainembodiments, the crystalline form loses about 0.1% of its total masswhen heated from about ambient temperature to about 220° C. In certainembodiments, the crystalline form is an anhydrate of Compound 1 andcorresponds to Form B.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 10 comprising an endothermicevent with on onset temperature at about 174° C. and a peak maximumtemperature at about 182° C. when heated from approximately 25° C. toapproximately 220° C.

In still another embodiment, Form B is substantially pure. In certainembodiments, the substantially pure Form B is substantially free ofother solid forms, e.g., amorphous solid. In another embodiment, Form Bis substantially free of Form A. In certain embodiments, the purity ofthe substantially pure Form B is no less than about 95%, no less thanabout 96%, no less than about 97%, no less than about 98%, no less thanabout 98.5%, no less than about 99%, no less than about 99.5%, or noless than about 99.8%.

Form C

In certain embodiments, provided herein is Form C.

In one embodiment, Form C is a solid form of Compound 1. In anotherembodiment, Form C is crystalline. In one embodiment, Form C is asolvated form of Compound 1. In one embodiment, Form C is anacetonitrile (MeCN) solvated form of Compound 1.

In certain embodiments, Form C provided herein is obtained byequilibration experiments, evaporation experiments, coolingrecrystallization experiments and anti-solvent recrystallizationexperiments (see Table 6, Table 7, and Table 9). In certain embodiments,Form C is obtained from certain solvent systems including MeCN orMeCN/H₂O (about 1:1). In certain embodiments, Form C is obtained fromcertain solvent systems including MeCN or MeCN/H₂O (about 1:1) at atemperature of about 50° C. In another embodiment, Form C is obtainedfrom a solution of 2-MeTHF/H₂O (about 1:1) distilled under vacuum atconstant volume with addition of MeCN.

In certain embodiments, a solid form provided herein, e.g., Form C, issubstantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, Form C has an X-ray powderdiffraction pattern substantially as shown in FIG. 11. In oneembodiment, a solid form provided herein, e.g. Form C, has one or morecharacteristic X-ray powder diffraction peaks at approximately 3.1, 7.7,8.9, 10.3, 13.3, 13.7, 14.2, 14.6, 14.8, 15.0, 15.3, 15.5, 15.7, 16.9,17.4, 17.8, 18.3, 18.7, 19.5, 19.9, 20.7, 21.1, 21.4, 22.1, 22.4, 22.7,23.1, 23.9, 24.6, 25.0, 25.5, 25.8, 26.1, 26.7, 26.9, 27.2, 27.7, 28.5,29.4, 29.8, 30.3, 30.9, 31.3, 32.4, 33.0, 33.6, 34.3, 35.4, 35.9, 36.2,37.1, 37.9, or 38.9° 2θ (±0.2° 2θ) or (±0.1° 2θ) as depicted in FIG. 11.In a specific embodiment, a solid form provided herein has one, two,three, four, five, six, seven, eight, or nine characteristic X-raypowder diffraction peaks at approximately 7.7, 8.9, 10.3, 15.3, 17.4,18.3, 19.9, 21.4, or 28.5° 2θ (±0.2° 2θ). In one embodiment, the solidform is Form C. In another embodiment, a solid form provided herein,e.g. Form C, has one, two, three or four characteristic X-ray powderdiffraction peaks at approximately 7.7, 8.9, 10.3, or 18.3° 2θ (±0.2°2θ). In another embodiment, Form C has one, two, three, four, five, six,seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen,sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two,twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven,twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three,thirty-four, thirty-five, thirty-six, thirty-seven, thirty-eight,thirty-nine, forty, forty-one, forty-two, forty-three, forty-four,forty-five, forty-six, forty-seven, forty-eight, forty-nine, fifty,fifty-one, fifty-two, or fifty-three characteristic X-ray powderdiffraction peaks as set forth in Table 14.

In one embodiment, provided herein is a crystalline form of Compound 1having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 12. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 6.6% of the total mass of the samplebetween approximately 50° C. and approximately 175° C. when heated fromapproximately 25° C. to approximately 220° C. Thus, in certainembodiments, the crystalline form loses about 6.6% of its total masswhen heated from about ambient temperature to about 220° C. Thetheoretical MeCN content of MeCN mono-solvate of Compound 1 is 6.7% byweight, matching the TGA weight loss observed.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 13 comprising an endothermicevent with a maximum at about 165° C. when heated from approximately 25°C. to approximately 300° C. In another embodiment, provided herein is acrystalline form of Compound 1 having a DSC thermogram as depicted inFIG. 13 further comprising an endothermic event with a maximum at about186° C. when heated from approximately 25° C. to approximately 300° C.

In one embodiment, provided herein is a solid form, e.g. Form C, havinga ¹H NMR spectrum substantially as depicted in FIG. 14.

In still another embodiment, Form C is substantially pure. In certainembodiments, the substantially pure Form C is substantially free ofother solid forms, e.g., amorphous solid. In certain embodiments, thepurity of the substantially pure Form C is no less than about 95%, noless than about 96%, no less than about 97%, no less than about 98%, noless than about 98.5%, no less than about 99%, no less than about 99.5%,or no less than about 99.8%.

Form D

In certain embodiments, provided herein is Form D.

In one embodiment, Form D is a solid form of Compound 1. In anotherembodiment, Form D is crystalline. In one embodiment, Form D is asolvated form of Compound 1. In one embodiment, Form D is an IPAsolvated form of Compound 1.

In certain embodiments, Form D provided herein is obtained byequilibration experiments, evaporation experiments, coolingrecrystallization experiments and anti-solvent recrystallizationexperiments (see Table 6, Table 7, and Table 9). In certain embodiments,Form D is obtained from certain solvent systems including IPA at roomtemperature.

In certain embodiments, a solid form provided herein, e.g., Form D, issubstantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, Form D has an X-ray powderdiffraction pattern substantially as shown in FIG. 15. In oneembodiment, a solid form provided herein, e.g. Form D, has one or morecharacteristic X-ray powder diffraction peaks at approximately 3.1, 5.9,7.4, 8.7, 10.1, 11.1, 13.7, 14.8, 15.1, 16.3, 16.6, 17.6, 18.1, 19.2,19.8, 20.4, 21.5, 22.1, 22.3, 24.0, 24.3, 25.0, 26.2, 26.9, 27.3, 27.6,28.2, 28.6, 30.9, 31.4, 32.9, 33.6, 34.6, or 37.2° 2θ (±0.2° 2θ) or(±0.1° 2θ) as depicted in FIG. 15. In a specific embodiment, a solidform provided herein has one, two, three, or four characteristic X-raypowder diffraction peaks at approximately 7.4, 8.7, 10.1, or 18.1° 2θ(±0.2° 2θ). In one embodiment, the solid form is Form D. In anotherembodiment, Form D has one, two, three, four, five, six, seven, eight,nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen,seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two,twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven,twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three,or thirty-four characteristic X-ray powder diffraction peaks as setforth in Table 15.

In one embodiment, a solid form provided herein, e.g. Form D has a SEMimage substantially as shown in FIG. 16.

In one embodiment, provided herein is a crystalline form of Compound 1having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 17. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 7.4% of the total mass of the samplebetween approximately 100° C. and approximately 160° C. when heated fromapproximately 25° C. to approximately 220° C. Thus, in certainembodiments, the crystalline form loses about 7.4% of its total masswhen heated from about ambient temperature to about 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 18 comprising an endothermicevent with an onset temperature at about 125° C. and a peak maximumtemperature at about 154° C. when heated from approximately 25° C. toapproximately 220° C. In one embodiment, provided herein is acrystalline form of Compound 1 having a DSC thermogram as depicted inFIG. 18 further comprising an endothermic event with an onsettemperature at about 175° C. and a peak maximum temperature at about185° C. when heated from approximately 25° C. to approximately 220° C.

In one embodiment, provided herein is a solid form provided herein, e.g.Form D, having a ¹H NMR spectrum substantially as depicted in FIG. 19.

In still another embodiment, Form D is substantially pure. In certainembodiments, the substantially pure Form D is substantially free ofother solid forms, e.g., amorphous solid. In certain embodiments, thepurity of the substantially pure Form D is no less than about 95%, noless than about 96%, no less than about 97%, no less than about 98%, noless than about 98.5%, no less than about 99%, no less than about 99.5%,or no less than about 99.8%.

Form E

In certain embodiments, provided herein is Form E.

In one embodiment, Form E is a solid form of Compound 1. In anotherembodiment, Form E is crystalline. In one embodiment, Form E is asolvated form of Compound 1. Form E can be an ethanol solvate where thesolvate optionally contains water.

In certain embodiments, Form E provided herein is obtained byequilibration experiments and evaporation experiments (see Table 6,Table 7, and Table 9). In certain embodiments, Form E is obtained fromcertain solvent systems including EtOH or EtOH/water (about 1:1). Incertain embodiments, Form E is obtained from certain solvent systemsincluding EtOH or EtOH/water (about 1:1) at a temperature of about 50°C.

In certain embodiments, a solid form provided herein, e.g., Form E, issubstantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, Form E has an X-ray powderdiffraction pattern substantially as shown in FIG. 20. In oneembodiment, a solid form provided herein, e.g. Form E, has one or morecharacteristic X-ray powder diffraction peaks at approximately 3.1, 5.5,7.8, 11.0, 13.5, 14.6, 15.6, 16.6, 17.5, 18.4, 20.0, 20.7, 22.2, 22.9,23.5, 24.2, 24.8, 26.1, 26.7, 27.3, 27.8, 28.4, 29.5, 30.0, 31.1, 31.6,32.1, 32.6, 33.6, 34.0, 34.5, 35.4, 36.3, 37.2, 38.1, 39.4, or 39.8° 2θ(±0.2° 2θ) or (±0.1° 2θ) as depicted in FIG. 20. In a specificembodiment, a solid form provided herein has one, two, three, four,five, six, or seven characteristic X-ray powder diffraction peaks atapproximately 7.8, 14.6, 15.6, 17.5, 22.2, 23.5, or 26.1° 2θ (±0.2° 2θ).In one embodiment, the solid form is Form E. In another embodiment, asolid form provided herein, e.g. Form E, has one, two, three or fourcharacteristic X-ray powder diffraction peaks at approximately 7.8,14.6, 17.5, or 22.2° 2θ (±0.2° 20). In another embodiment, Form E hasone, two, three, four, five, six, seven, eight, nine, ten, eleven,twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen,nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four,twenty-five, twenty-six, twenty-seven, twenty-eight, twenty-nine,thirty, thirty-one, thirty-two, thirty-three, thirty-four, thirty-five,thirty-six, or thirty-seven characteristic X-ray powder diffractionpeaks as set forth in Table 16.

In one embodiment, provided herein is a crystalline form of Compound 1having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 21. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 13.7% of the total mass of the samplebetween approximately 35° C. and approximately 175° C. when heated fromapproximately 35° C. to approximately 220° C. Thus, in certainembodiments, the crystalline form loses about 13.7% of its total masswhen heated from about ambient temperature to about 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 22 comprising a broadendothermic event with and onset temperature at about 92° C. and amaximum at about 104° C. when heated from approximately 25° C. toapproximately 220° C.

In still another embodiment, Form E is substantially pure. In certainembodiments, the substantially pure Form E is substantially free ofother solid forms, e.g., amorphous solid. In certain embodiments, thepurity of the substantially pure Form E is no less than about 95%, noless than about 96%, no less than about 97%, no less than about 98%, noless than about 98.5%, no less than about 99%, no less than about 99.5%,or no less than about 99.8%.

Form F

In certain embodiments, provided herein is Form F.

In one embodiment, Form F is a solid form of Compound 1. In anotherembodiment, Form F is crystalline. In one embodiment, Form F is asolvated form of Compound 1. In one embodiment, Form F is an IPAsolvated form of Compound 1.

In certain embodiments, Form F provided herein is obtained byequilibration experiments (see Table 6, Table 7, and Table 9). Incertain embodiments, Form F is obtained from certain solvent systemsincluding IPA or IPA/water at about 50° C.

In certain embodiments, a solid form provided herein, e.g., Form F, issubstantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, Form F has an X-ray powderdiffraction pattern substantially as shown in FIG. 23. In oneembodiment, a solid form provided herein, e.g. Form F, has one or morecharacteristic X-ray powder diffraction peaks at approximately 4.9, 7.0,9.4, 11.1, 11.8, 15.5, 15.8, 17.0, 17.6, 18.0, 18.3, 19.0, 19.7, 20.0,20.3, 20.9, 22.4, 22.6, 23.2, 23.7, 24.4, 25.1, 25.4, 25.6, 26.4, 26.8,27.3, 27.7, 28.6, 29.2, 30.0, 30.4, 30.7, 31.2, 32.1, 34.1, 34.4, 35.2,35.8, 36.5, 38.5, 38.8, or 39.2° 2θ (±0.2° 2θ) or (±0.1° 2θ) as depictedin FIG. 23. In a specific embodiment, a solid form provided herein, e.g.Form F, has one, two, three, four, five, six, seven, eight, or ninecharacteristic X-ray powder diffraction peaks at approximately 7.0, 9.4,11.8, 15.5, 18.0, 18.3, 19.7, 20.0, or 20.9° 2θ (±0.2° 2θ). In anotherembodiment, a solid form provided herein has one, two, three or fourcharacteristic X-ray powder diffraction peaks at approximately 9.4,11.8, 18.0, or 18.3° 2θ (±0.2° 2θ). In one embodiment, the solid form isForm F. In another embodiment, Form F has one, two, three, four, five,six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen,fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one,twenty-two, twenty-three, twenty-four, twenty-five, twenty-six,twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two,thirty-three, thirty-four, thirty-five, thirty-six, thirty-seven,thirty-eight, thirty-nine, forty, forty-one, forty-two, or forty-threecharacteristic X-ray powder diffraction peaks as set forth in Table 17.

In one embodiment, a solid form provided herein, e.g. Form F has a SEMimage substantially as shown in FIG. 24.

In one embodiment, provided herein is a crystalline form of Compound 1having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 25. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 14.3% of the total mass of the samplebetween approximately 50° C. and approximately 175° C. when heated fromapproximately 50° C. to approximately 220° C. Thus, in certainembodiments, the crystalline form loses about 14.3% of its total masswhen heated from about ambient temperature to about 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 26 comprising an endothermicevent with an onset temperature at about 137° C. and a peak maximumtemperature at about 152° C. when heated from approximately 25° C. toapproximately 220° C.

In one embodiment, provided herein is a solid form provided herein, e.g.Form F, having a ¹H NMR spectrum substantially as depicted in FIG. 27.

In still another embodiment, Form F is substantially pure. In certainembodiments, the substantially pure Form F is substantially free ofother solid forms, e.g., amorphous solid. In certain embodiments, thepurity of the substantially pure Form F is no less than about 95%, noless than about 96%, no less than about 97%, no less than about 98%, noless than about 98.5%, no less than about 99%, no less than about 99.5%,or no less than about 99.8%.

Form G

In certain embodiments, provided herein is Form G.

In one embodiment, Form G is a solid form of Compound 1. In anotherembodiment, Form G is crystalline. In one embodiment, Form G is asolvated form of Compound 1. In one embodiment, Form G is a MTBEsolvated form of Compound 1.

In certain embodiments, Form G provided herein is obtained byequilibration experiments, evaporation experiments and anti-solventrecrystallization experiments (see Table 6, Table 7, and Table 9). Incertain embodiments, Form G is obtained from certain solvent systemsincluding MTBE. In certain embodiments, Form G is obtained from certainsolvent systems including MTBE at a temperature of 50° C.

In certain embodiments, a solid form provided herein, e.g., Form G, issubstantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, Form G has an X-ray powderdiffraction pattern substantially as shown in FIG. 28. In oneembodiment, Form G has one or more characteristic X-ray powderdiffraction peaks at approximately 4.5, 8.0, 9.0, 9.9, 10.0, 10.2, 11.6,11.9, 13.5, 14.4, 14.6, 15.3, 15.9, 16.4, 16.9, 17.5, 17.7, 18.0, 18.4,18.7, 18.8, 19.4, 19.6, 20.3, 20.8, 21.2, 21.6, 22.0, 22.2, 22.5, 22.9,23.4, 24.0, 24.5, 24.6, 25.0, 25.2, 25.6, 25.9, 26.0, 26.4, 26.9, 27.3,27.6, 28.0, 28.2, 28.8, 29.4, 29.9, 30.2, 30.8, 31.4, 31.8, 32.8, 33.2,34.4, 34.9, 35.7, 36.1, 38.2, or 38.9° 2θ (±0.2° 2θ) or (±0.1° 20). In aspecific embodiment, a solid form provided herein, e.g. Form G, has one,two, three, four, five, six, seven, eight, nine, ten, or elevencharacteristic X-ray powder diffraction peaks at approximately 9.0, 9.9,10.0, 15.3, 17.5, 18.4, 18.7, 19.4, 19.6, 21.2, or 22.9° 2θ (±0.2° 2θ).In another embodiment, a solid form provided herein has one, two, three,or four characteristic X-ray powder diffraction peaks at approximately9.9, 15.3, 18.4, or 22.9° 2θ (±0.2° 2θ). In one embodiment, the solidform is Form G. In another embodiment, Form G has one, two, three, four,five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen,fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one,twenty-two, twenty-three, twenty-four, twenty-five, twenty-six,twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two,thirty-three, thirty-four, thirty-five, thirty-six, thirty-seven,thirty-eight, thirty-nine, forty, forty-one, forty-two, forty-three,forty-four, forty-five, forty-six, forty-seven, forty-eight, forty-nine,fifty, fifty-one, fifty-two, fifty-three, fifty-four, fifty-five,fifty-six, fifty-seven, fifty-eight, fifty-nine, sixty, or sixty-onecharacteristic X-ray powder diffraction peaks as set forth in Table 18.

In one embodiment, Form G has a SEM image substantially as shown in FIG.29.

In one embodiment, provided herein is a crystalline form of Compound 1having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 30. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 1.1% of the total mass of the samplebetween approximately 50° C. and approximately 140° C. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 8.7% of the total mass of the samplebetween approximately 50° C. and approximately 180° C. when heated fromapproximately 25° C. to approximately 220° C. Thus, in certainembodiments, the crystalline form loses about 8.7% of its total masswhen heated from about ambient temperature to about 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 31 comprising an endothermicevent with an onset temperature at about 144° C. and a peak maximumtemperature at about 148° C. when heated from approximately 25° C. toapproximately 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 31 comprising an endothermicevent with a maximum at about 161° C. when heated from approximately 25°C. to approximately 220° C.

In one embodiment, provided herein is a solid form provided herein, e.g.Form G, having a ¹H NMR spectrum substantially as depicted in FIG. 32.In one embodiment, the ¹H NMR spectrum of Form G shows Form G contains asignificant amount of MBTE.

In still another embodiment, Form G is substantially pure. In certainembodiments, the substantially pure Form G is substantially free ofother solid forms, e.g., amorphous solid. In certain embodiments, thepurity of the substantially pure Form G is no less than about 95%, noless than about 96%, no less than about 97%, no less than about 98%, noless than about 98.5%, no less than about 99%, no less than about 99.5%,or no less than about 99.8%.

Form H

In certain embodiments, provided herein is Form H.

In one embodiment, Form H is a solid form of Compound 1. In anotherembodiment, Form H is crystalline. In one embodiment, Form H is asolvated form of Compound 1. In one embodiment, Form H is an EtOHsolvated form of Compound 1. In certain embodiments, Form H can beconverted to Form A by contact with an environment comprising at least20% RH.

In certain embodiments, Form H provided herein is obtained byequilibration experiments, evaporation experiments and anti-solventrecrystallization experiments (see Table 6, Table 7, and Table 9). Incertain embodiments, Form H is obtained from certain solvent systemsincluding EtOH, EtOH/water (about 1:1), or EtOAc. In certainembodiments, Form H is obtained from certain solvent systems includingEtOH, EtOH/water (about 1:1), or EtOAc at a temperature of 50° C.

In certain embodiments, a solid form provided herein, e.g., Form H, issubstantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, Form H has an X-ray powderdiffraction pattern substantially as shown in FIG. 33. In oneembodiment, a solid form provided herein, e.g. Form H, has one or morecharacteristic X-ray powder diffraction peaks at approximately 6.1, 7.7,8.9, 10.3, 10.9, 11.3, 11.6, 13.7, 14.4, 15.0, 15.2, 15.4, 15.6, 15.9,16.9, 17.2, 17.7, 18.2, 18.7, 19.4, 19.6, 20.6, 20.9, 21.4, 22.5, 23.2,23.7, 24.6, 24.9, 25.6, 25.9, 26.2, 26.9, 27.4, 28.1, 28.4, 29.0, 29.4,31.1, 32.2, 33.1, 34.1, 34.7, 35.3, 37.3, or 38.6° 2θ (±0.2° 2θ) or(±0.1° 2θ) as depicted in FIG. 33. In a specific embodiment, a solidform provided herein, e.g. Form H, has one, two, three, four, five, six,seven, eight, nine, ten, or eleven characteristic X-ray powderdiffraction peaks at approximately 7.7, 8.9, 10.3, 15.2, 15.4, 15.6,17.2, 18.2, 19.6, 21.4, or 24.9° 2θ (±0.2° 2θ). In another embodiment, asolid form provided herein has one, two, three, or four characteristicX-ray powder diffraction peaks at approximately 7.7, 8.9, 10.3, or 18.2°2θ (±0.2° 2θ). In one embodiment, the solid form is Form H. In anotherembodiment, Form H has one, two, three, four, five, six, seven, eight,nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen,seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two,twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven,twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three,thirty-four, thirty-five, thirty-six, thirty-seven, thirty-eight,thirty-nine, forty, forty-one, forty-two, forty-three, forty-four,forty-five, or forty-six characteristic X-ray powder diffraction peaksas set forth in Table 19.

In one embodiment, provided herein is a crystalline form of Compound 1having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 34. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 6.5% of the total mass of the samplebetween approximately 50° C. and approximately 175° C. when heated fromapproximately 50° C. to approximately 220° C. Thus, in certainembodiments, the crystalline form loses about 6.5% of its total masswhen heated from about ambient temperature to about 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 35 comprising an endothermicevent with a peak maximum at about 163° C. when heated fromapproximately 25° C. to approximately 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 35 comprising an endothermicevent with an onset temperature at about 179° C. and a peak maximumtemperature at about 187° C. when heated from approximately 25° C. toapproximately 220° C.

In still another embodiment, Form H is substantially pure. In certainembodiments, the substantially pure Form H is substantially free ofother solid forms, e.g., amorphous solid. In certain embodiments, thepurity of the substantially pure Form H is no less than about 95%, noless than about 96%, no less than about 97%, no less than about 98%, noless than about 98.5%, no less than about 99%, no less than about 99.5%,or no less than about 99.8%.

Form I

In certain embodiments, provided herein is Form I.

In one embodiment, Form I is a solid form of Compound 1. In anotherembodiment, Form I is crystalline. In one embodiment, Form I is asolvated form of Compound 1. In one embodiment, Form I is a MeCNsolvated form of Compound 1.

In certain embodiments, Form I provided herein is obtained by coolingrecrystallization experiments and anti-solvent recrystallizationexperiments. In certain embodiments, Form I is obtained from certainsolvent systems including MeCN. In certain embodiments, Form I canconvert to Form C in a MeCN slurry at room temperature.

In certain embodiments, a solid form provided herein, e.g., Form I, andis substantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, Form I has an X-ray powderdiffraction pattern substantially as shown in FIG. 36. In oneembodiment, a solid form provided herein, e.g. Form I, has one or morecharacteristic X-ray powder diffraction peaks at approximately 5.2, 5.5,6.3, 8.6, 9.3, 10.4, 10.9, 11.5, 11.9, 12.6, 15.7, 16.6, 17.3, 18.1,18.7, 19.0, 20.0, 20.9, 22.0, 22.5, 23.3, 24.1, 24.6, 25.4, 26.4, 27.6,28.4, 29.6, 31.0, 31.6, 32.1, 33.2, 33.9, 35.3, 35.9, or 38.5° 2θ (±0.2°2θ) or (±0.1° 2θ) as depicted in FIG. 36. In a specific embodiment, asolid form provided herein has one, two, three, or four characteristicX-ray powder diffraction peaks at approximately 6.3, 15.7, 18.1, or20.0° 2θ (±0.2° 20). In one embodiment, the solid form is Form I. Inanother embodiment, Form I has one, two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen,seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two,twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven,twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three,thirty-four, thirty-five, or thirty-six characteristic X-ray powderdiffraction peaks as set forth in Table 20.

In one embodiment, provided herein is a crystalline form of Compound 1having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 38. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 2.3% of the total mass of the samplebetween approximately 50° C. and approximately 180° C. when heated fromapproximately 50° C. to approximately 220° C. Thus, in certainembodiments, the crystalline form loses about 2.3% of its total masswhen heated from about ambient temperature to about 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 39 comprising an endothermicevent with a peak maximum at about 75° C. when heated from approximately25° C. to approximately 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 39 comprising an endothermicevent with an onset temperature of about 173° C. and a peak maximumtemperature at about 183° C. when heated from approximately 25° C. toapproximately 220° C.

In one embodiment, provided herein is a solid form provided herein, e.g.Form I, having a ¹H NMR spectrum substantially as depicted in FIG. 37.

In still another embodiment, Form I is substantially pure. In certainembodiments, the substantially pure Form I is substantially free ofother solid forms, e.g., amorphous solid. In certain embodiments, thepurity of the substantially pure Form I is no less than about 95%, noless than about 96%, no less than about 97%, no less than about 98%, noless than about 98.5%, no less than about 99%, no less than about 99.5%,or no less than about 99.8%.

Amorphous Solid

In certain embodiments, provided herein is an amorphous solid ofCompound 1.

In certain embodiments, the amorphous solid provided herein is obtainedby evaporation and/or heat treatment of Form A.

In one embodiment, the amorphous solid has an X-ray powder diffractionspectrum substantially as shown in FIG. 40.

In one embodiment, provided herein is an amorphous solid of Compound 1having a DSC thermogram as depicted in FIG. 41 comprising a glasstransition temperature of 84° C. when heated from approximately 40° C.to approximately 260° C.

In one embodiment, provided herein is an amorphous solid of Compound 1having a ¹H NMR spectrum substantially as depicted in FIG. 42.

In one embodiment, provided herein is an amorphous solid of Compound 1having a DVS isotherm plot substantially as depicted in FIG. 43A.

In still another embodiment, the amorphous solid of Compound 1 issubstantially pure. In certain embodiments, the substantially pureamorphous solid of Compound 1 is substantially free of other solidforms, e.g., Form A, Form B, Form C, Form D, Form E, Form F, Form G,Form H, and Form I. In certain embodiments, the purity of thesubstantially pure amorphous solid is no less than about 95%, no lessthan about 96%, no less than about 97%, no less than about 98%, no lessthan about 98.5%, no less than about 99%, no less than about 99.5%, orno less than about 99.8%.

Citrate Salt Form Y

Also provided herein are solid forms of Compound 1 that include citratesalts.

In certain embodiments, provided herein is citrate salt Form Y.

In one embodiment, the citrate salt Form Y is a solid form ofCompound 1. In another embodiment, the citrate salt Form Y iscrystalline. In another embodiment, the citrate salt Form Y is ananhydrate.

In certain embodiments, the citrate salt Form Y provided herein isobtained by equilibration experiments, evaporation experiments andanti-solvent recrystallization experiments (see Table 23, Table 24, andTable 25). In certain embodiments, the citrate salt Form Y is obtainedfrom certain solvent systems including acetone, MeCN, n-butanol, EtOH,EtOH/water (about 1:1), EtOAc, heptanes, IPA, DCM, MeOAc, MTBE, MEK,toluene, THF, THF/water (about 1:1), 1,4-dioxane, MIBK, IPAc, and2-MeTHF. In certain embodiments, the citrate salt Form Y is obtainedfrom certain solvent systems including acetone, MeCN, n-butanol, EtOH,EtOH/water (about 1:1), EtOAc, heptanes, IPA, DCM, MeOAc, MTBE, MEK,toluene, THF, THF/water (about 1:1), 1,4-dioxane, MIBK, IPAc, and2-MeTHF at 50° C. In one embodiment, the citrate salt Form Y is an EtOHsolvate.

In one embodiment, a method of preparing the citrate salt Form Ycomprises the steps of cooling to a temperature less than about 50° C.in THF or THF/water and collecting solids.

In certain embodiments, a solid form provided herein, e.g., Form Y is acitrate salt of Compound 1, and is substantially crystalline, asindicated by, e.g., X-ray powder diffraction measurements. In oneembodiment, a solid form provided herein, e.g., Form Y, has an X-raypowder diffraction pattern (XRPD) substantially as shown in FIG. 45. Inone embodiment, a solid form provided herein, e.g., Form Y, has one ormore characteristic X-ray powder diffraction peaks at approximately 4.8,6.6, 9.6, 13.6, 14.4, 15.4, 16.0, 16.9, 18.0, 18.9, 19.2, 19.9, 20.1,20.9, 21.8, 22.4, 22.7, 23.2, 23.4, 24.0, 24.1, 24.3, 25.1, 26.7, 27.0,27.9, 28.5, 29.0, 29.6, 30.2, 30.4, 30.8, 31.1, 31.6, 32.3, 33.1, 33.5,34.0, 34.6, or 35.1° 2θ (±0.2° 2θ) or (±0.1° 20) as depicted in FIG. 45.In a specific embodiment, a solid form provided herein, e.g., Form Y,has one, two, three, four, five, six, seven, eight, nine, ten, or elevencharacteristic X-ray powder diffraction peaks at approximately 4.8, 6.6,9.6, 15.4, 16.0, 16.9, 18.9, 19.2, 19.9, 20.9, or 28.5° 2θ (±0.2° 2θ).In another embodiment, a solid form provided herein has one, two, three,or four characteristic X-ray powder diffraction peaks at approximately4.8, 9.6, 18.9, or 19.2° 2θ (±0.2° 20). In one embodiment, the solidform is citrate salt Form Y. In another embodiment, the citrate saltForm Y has one, two, three, four, five, six, seven, eight, nine, ten,eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen,eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three,twenty-four, twenty-five, twenty-six, twenty-seven, twenty-eight,twenty-nine, thirty, thirty-one, thirty-two, thirty-three, thirty-four,thirty-five, thirty-six, thirty-seven, thirty-eight, thirty-nine, orforty characteristic X-ray powder diffraction peaks as set forth inTable 27.

In one embodiment, a solid form provided herein, e.g., Form Y, has a SEMimage substantially as shown in FIG. 46.

In one embodiment, provided herein is a crystalline citrate salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 47. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 0.1% of the total mass of the samplebetween approximately 50° C. and approximately 150° C. when heated fromapproximately 50° C. to approximately 220° C.

In one embodiment, provided herein is a crystalline citrate salt ofCompound 1 having a DSC thermogram substantially as depicted in FIG. 48comprising an endothermic event with an onset temperature of about 213°C. and a peak maximum temperature at about 217° C. when heated fromapproximately 25° C. to approximately 260° C.

In one embodiment, provided herein is a solid form, e.g., Form Y, havinga DVS isotherm plot substantially as depicted in FIG. 49A.

In one embodiment, provided herein a solid form, e.g., Form Y, having a¹H NMR spectrum substantially as depicted in FIG. 50.

In still another embodiment, the citrate salt Form Y is substantiallypure. In certain embodiments, the substantially pure citrate salt Form Yis substantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure citrate saltForm Y is no less than about 95%, no less than about 96%, no less thanabout 97%, no less than about 98%, no less than about 98.5%, no lessthan about 99%, no less than about 99.5%, or no less than about 99.8%.

Citrate Salt Form Z

In certain embodiments, provided herein is a citrate salt Form Z.

In one embodiment, the citrate salt Form Z is a solid form ofCompound 1. In another embodiment, the citrate salt Form Z iscrystalline. In another embodiment, the citrate salt Form Z is ananhydrate. In another embodiment, the citrate salt Form Z is a hydrate.In one embodiment, the citrate salt Form Z is a non-stoichiometrichydrate. In still another embodiment, the citrate salt Form Z is achannel hydrate. In still another embodiment, the citrate salt Form Z isa non-stoichiometric channel hydrate. In still another embodiment, thecitrate salt Form Z is a solvate.

In certain embodiments, Form Z is obtained by equilibration experiments,evaporation experiments and anti-solvent recrystallization experiments(see Table 23, Table 24, and Table 25). In certain embodiments, thecitrate salt Form Z is obtained from certain solvent systems includingMeCN/water (about 1:1), EtOH, EtOH/water (about 1:1), or MeOH. Incertain embodiments, the citrate salt Form Z is obtained from certainsolvent systems including MeCN/water (about 1:1), EtOH, EtOH/water(about 1:1), or MeOH at a temperature of about 50° C.

In one embodiment, a solid form provided herein, e.g., Form Z, has a SEMimage substantially as shown in FIG. 53.

In certain embodiments, a solid form provided herein, e.g., Form Z, issubstantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, a solid form providedherein, e.g., Form Z, has an X-ray powder diffraction patternsubstantially as shown in FIG. 52. In one embodiment, a solid formprovided herein, e.g., Form Z, has one or more characteristic X-raypowder diffraction peaks at approximately 4.6, 6.6, 9.4, 13.1, 14.1,15.3, 15.6, 17.4, 18.8, 19.0, 19.9, 20.4, 21.1, 21.9, 22.2, 22.7, 23.5,23.9, 25.2, 26.3, 26.8, 27.8, 28.3, 28.7, 29.8, 31.2, 31.9, 32.6, 33.7,35.1, 35.9, 37.4, or 38.0° 2θ (±0.2° 2θ) or (±0.1° 2θ) as depicted inFIG. 52. In a specific embodiment, a solid form provided herein has one,two, three, or four characteristic X-ray powder diffraction peaks atapproximately 9.4, 18.8, 19.0, or 28.7° 2θ (±0.2° 2θ). In oneembodiment, the solid form is citrate salt Form Z. In anotherembodiment, the citrate salt Form Z has one, two, three, four, five,six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen,fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one,twenty-two, twenty-three, twenty-four, twenty-five, twenty-six,twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two,or thirty-three characteristic X-ray powder diffraction peaks as setforth in Table 30.

In one embodiment, provided herein is a crystalline citrate salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 54. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 0.1% of the total mass of the samplebetween approximately 50° C. and approximately 150° C. when heated fromapproximately 25° C. to approximately 300° C. In certain embodiments,the crystalline form is an anhydrate of Compound 1.

In one embodiment, provided herein is a crystalline citrate salt Form ofCompound 1 having a DSC thermogram as depicted in FIG. 55 comprising anendothermic event with an onset temperature at about 217° C. and a peakmaximum temperature at about 221° C. when heated from approximately 25°C. to approximately 260° C.

In one embodiment, provided herein is a solid form, e.g., Form Y, havinga DVS isotherm plot substantially as depicted in FIG. 56A.

In one embodiment, provided herein is a solid form provided herein,e.g., Form Z, having a ¹H NMR spectrum substantially as depicted in FIG.57.

In one embodiment, provided herein is a hydrate of the citrate salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 58. In certainembodiments, the hydrate exhibits a TGA thermogram comprising a totalmass loss of approximately 2% of the total mass of the sample betweenapproximately 25° C. and approximately 200° C. when heated fromapproximately 25° C. to approximately 300° C. In certain embodiments,the crystalline form is a hydrate of the Citrate form of Compound 1.

In one embodiment, provided herein is a non-stoichiometric hydrate ofthe citrate salt of Compound 1 having a TGA thermograph correspondingsubstantially to the representative TGA thermogram as depicted in FIG.59. In certain embodiments, the non-stoichiometric hydrate form exhibitsa TGA thermogram comprising a total mass loss of approximately 1.7% ofthe total mass of the sample between approximately 50° C. andapproximately 200° C. when heated from approximately 50° C. toapproximately 300° C. In certain embodiments, the crystalline form is anon-stoichiometric hydrate of the Citrate form of Compound 1.

In one embodiment, provided herein is a solvate of the citrate salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 60. In certainembodiments, the solvate exhibits a TGA thermogram comprising a totalmass loss of approximately 1.3% of the total mass of the sample betweenapproximately 25° C. and approximately 200° C. when heated fromapproximately 25° C. to approximately 300° C. In certain embodiments,the crystalline form is a solvate of the Citrate form of Compound 1.

In one embodiment, provided herein is a solid form provided herein,e.g., solvate of Form Z, having a ¹H NMR spectrum substantially asdepicted in FIG. 61.

In still another embodiment, the citrate salt Form Z is substantiallypure. In certain embodiments, the substantially pure citrate salt Form Zis substantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure citrate saltForm Z is no less than about 95%, no less than about 96%, no less thanabout 97%, no less than about 98%, no less than about 98.5%, no lessthan about 99%, no less than about 99.5%, or no less than about 99.8%.

HCl Salt Forms

In certain embodiments provided herein is a starting material HCl saltForm. In one embodiment, the starting material HCl salt Form is a solidform of Compound 1. In another embodiment, the starting material HClsalt Form is an anhydrate.

In one embodiment, is the starting material HCl salt Form is obtained bydissolving Compound 1 in MeOH (e.g., about 10 Vol.) at a temperature ofabout 25° C. to about 30° C. HCl in MeOH (˜1.25 M, 1.10 eq) is added toobtain the HCl salt Form starting material of Compound 1. The solutioncan be vacuum distilled and the solvent changed from MeOH to EtOAc(e.g., about 30-35 Vol.), where the temperature is optionally maintainedat about 25° C. to about 35° C. The slurry can be filtered and the cakewashed with EtOAc (e.g., about 5 Vol.). The cake can be dried in avacuum oven at 50° C.

In certain embodiments, a solid form provided herein, e.g., a startingmaterial HCl salt Form, is substantially crystalline, as indicated by,e.g., X-ray powder diffraction measurements. In one embodiment, a solidform provided herein, e.g., a starting material HCl salt Form, has anX-ray powder diffraction pattern substantially as shown in FIG. 63. Inone embodiment, a solid form provided herein, e.g., a starting materialHCl salt Form, has one or more characteristic X-ray powder diffractionpeaks at approximately 5.8, 7.1, 8.3, 10.1, 11.3, 11.6, 12.7, 15.5,16.1, 17.8, 19.2, 19.7, 20.5, 21.1, 23.0, 24.0, 25.5, 26.3, 27.2, 28.4,31.0, or 35.6° 2θ (±0.2° 2θ) or (±0.1° 2θ) as depicted in FIG. 63. In aspecific embodiment, a solid form provided herein has one, two, three,or four characteristic X-ray powder diffraction peaks at approximately21.1, 19.2, 20.5, 17.8° 2θ (±0.2° 2θ) as depicted in FIG. 63. In oneembodiment, the solid form is a starting material HCl salt Form. Inanother embodiment, the starting material HCl salt Form has one, two,three, four, five, six, seven, eight, nine, ten, eleven, twelve,thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen,twenty, twenty-one, or twenty-two, characteristic X-ray powderdiffraction peaks as set forth in Table 42.

In one embodiment, provided herein is a starting material HCl salt Formof Compound 1 having a TGA thermograph corresponding substantially tothe representative TGA thermogram as depicted in FIG. 64. In certainembodiments, the starting material HCl salt Form exhibits a TGAthermogram comprising a total mass loss of approximately 1.1% of thetotal mass of the sample between approximately 24° C. and approximately100° C. when heated from approximately 24° C. to approximately 300° C.In certain embodiments, the crystalline form is an anhydrate of Compound1.

In one embodiment, provided herein is a starting material HCl salt Formof Compound 1 having a DSC thermogram as depicted in FIG. 64 comprisingan endothermic event with an onset temperature at about 239° C. and apeak maximum temperature at about 249° C. when heated from approximately24° C. to approximately 300° C.

In one embodiment, provided herein is a solid form, e.g., a startingmaterial HCl salt Form, having a DVS isotherm plot substantially asdepicted in FIG. 65.

In still another embodiment, the starting material HCl salt Form issubstantially pure. In certain embodiments, the substantially purestarting material HCl salt Form is substantially free of other solidforms, e.g., amorphous solid. In certain embodiments, the purity of thesubstantially pure starting material HCl salt Form is no less than about95%, no less than about 96%, no less than about 97%, no less than about98%, no less than about 98.5%, no less than about 99%, no less thanabout 99.5%, or no less than about 99.8%.

HCl Salt Form 1

In certain embodiments, provided herein is an HCl salt Form 1 ofCompound 1.

In one embodiment, the HCl salt Form 1 is a solid form of Compound 1. Inone embodiment, the HCl salt Form 1 is a solvate. In one embodiment, theHCl salt Form 1 is an IPA solvate form of Compound 1. In anotherembodiment, the HCl salt Form 1 is crystalline.

In certain embodiments, the HCl salt Form 1 provided herein is obtainedby equilibration experiments, evaporation experiments and anti-solventrecrystallization experiments (see see Table 23, Table 24, and Table25). In certain embodiments, the HCl salt Form 1 is obtained fromcertain solvent systems including IPA.

In one embodiment, a method of preparing the HCl salt Form 1 comprisesthe steps of dissolving Compound 1 in IPA and slowly evaporating the IPAand collecting solids.

In certain embodiments, a solid form provided herein, e.g., HCl saltForm 1, is an HCl salt of Compound 1, and is substantially crystalline,as indicated by, e.g., X-ray powder diffraction measurements. In oneembodiment, a solid form provided herein, e.g., HCl salt Form 1, has anX-ray powder diffraction pattern (XRPD) substantially as shown in FIG.66. In one embodiment, a solid form provided herein, e.g., HCl salt Form1, has one or more characteristic X-ray powder diffraction peaks atapproximately 5.5, 5.9, 5.9, 8.8, 9.4, 9.9, 11.5, 12.4, 12.8, 15.6,15.9, 16.2, 17.4, 18.6, 20.4, 20.7, 21.0, 21.3, 21.7, 22.1, 22.6, 22.8,22.9, 23.2, 23.5, 23.7, 23.9, 25.5, 25.8, 26.3, 26.8, 27.0, 28.4, 29.3,30.3, 32.1, 32.2, 33.5, 35.5, 36.6, or 37.6° 2θ (±0.2° 20) or (±0.1° 2θ)as depicted in FIG. 66. In a specific embodiment, a solid form providedherein has one, two, three, four, five, six, seven, eight, nine, ten,eleven, twelve, or thirteen characteristic X-ray powder diffractionpeaks at approximately 5.5 5.9, 8.8, 9.4, 15.9, 18.6, 20.7, 22.6, 22.8,22.9, 29.3, 32.1, or 32.2° 2θ (±0.2° 2θ). In one embodiment, the solidform is HCl Salt Form 1. In another embodiment, a solid form providedherein has one, two, three or four characteristic X-ray powderdiffraction peaks at approximately 8.8, 9.4, 15.9, or 20.7° 2θ (±0.2°20). In another embodiment, HCl Salt Form 1 has one, two, three, four,five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen,fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one,twenty-two, twenty-three, twenty-four, twenty-five, twenty-six,twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two,thirty-three, thirty-four, thirty-five, thirty-six, thirty-seven,thirty-eight, thirty-nine, forty, or forty-one characteristic X-raypowder diffraction peaks as set forth in Table 34.

In one embodiment, provided herein is crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 67. In certainembodiments, crystalline form HCl salt of Compound 1 exhibits a TGAthermogram comprising a total mass loss of approximately 6.6% of thetotal mass of the sample between approximately 20° C. and approximately140° C. when heated from approximately 20° C. to approximately 325° C.

In certain embodiments, a solid form provided herein exhibits a TGAthermogram comprising a total mass loss of approximately 10% of thetotal mass of the sample between approximately 150° C. and approximately200° C. when heated from approximately 20° C. to approximately 325° C.Thus, in certain embodiments, the crystalline form HCl salt of Compound1 loses about 17% of its total mass when heated from about ambienttemperature to about 325° C.

In one embodiment, provided herein is crystalline form HCl salt ofCompound 1 having a DSC thermogram substantially as depicted in FIG. 67comprising an endothermic event with an onset temperature at about 102°C. and a peak maximum temperature at about 114° C. when heated fromapproximately 25° C. to approximately 350° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram substantially as depicted in FIG. 67comprising an endothermic event with an onset temperature at about 146°C. and a peak maximum temperature at about 181° C. when heated fromapproximately 25° C. to approximately 350° C.

In still another embodiment, provided herein is a crystalline form HClsalt of Compound 1 having a DSC thermogram substantially as depicted inFIG. 67 comprising multiple endothermic events each having a maximumgreater than 250° C. when heated from approximately 25° C. toapproximately 350° C.

In still another embodiment, HCl salt Form 1 is substantially pure. Incertain embodiments, the substantially pure HCl salt Form 1 issubstantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure HCl salt Form1 is no less than about 95%, no less than about 96%, no less than about97%, no less than about 98%, no less than about 98.5%, no less thanabout 99%, no less than about 99.5%, or no less than about 99.8%.

HCl Salt Form 2

In certain embodiments, provided herein is the HCl salt Form 2.

In one embodiment, the HCl salt Form 2 is a solid form of Compound 1. Inanother embodiment, HCl Salt Form 2 is crystalline. In one embodiment,HCl Salt Form 2 is an anhydrate form of Compound 1. In one embodiment,HCl salt Form 2 is solvated form of Compound 1. In one embodiment, HClsalt Form 2 is an IPA solvated form of Compound 1. In one embodiment,HCl salt Form 2 is a toluene solvated form of Compound 1.

In certain embodiments, HCl salt Form 2 provided herein is obtained byequilibration experiments, evaporation experiments and anti-solventrecrystallization experiments. In certain embodiments, HCl salt Form 2is obtained from certain solvent systems including IPA/toluene (about1:1).

In certain embodiments, a solid form provided herein, e.g., HCl saltForm 2, is substantially crystalline, as indicated by, e.g., X-raypowder diffraction measurements. In one embodiment, HCl salt Form 2 hasan X-ray powder diffraction pattern substantially as shown in FIG. 68.In one embodiment, a solid form provided herein, e.g., HCl salt Form 2,has one or more characteristic X-ray powder diffraction peaks atapproximately 5.5, 5.8, 9.0, 9.8, 9.9, 10.7, 11.0, 12.3, 12.6, 14.0,16.7, 18.1, 19.0, 19.5, 19.9, 20.1, 20.8, 21.5, 21.8, 22.0, 22.8, 23.3,24.0, 24.3, 24.6, 24.9, 25.3, 26.0, 26.5, 26.8, 27.0, 27.6, 28.4, 29.2,29.7, 30.7, 33.0, or 35.1° 2θ (±0.2° 20) or (±0.1° 2θ) as depicted inFIG. 68. In a specific embodiment, a solid form provided herein, e.g.,HCl salt Form 2, has one, two, three, four, five, six, seven, eight,nine, ten, or eleven characteristic X-ray powder diffraction peaks atapproximately 5.5, 9.0, 9.8, 9.9, 12.3, 12.6, 16.7, 18.1, 19.0, 21.5, or22.8° 2θ (±0.2° 2θ). In another embodiment, a solid form provided hereinhas one, two, three, or four characteristic X-ray powder diffractionpeaks at approximately 9.0, 9.8, 9.9, or 16.7° 2θ (±0.2° 2θ). In oneembodiment, the solid form is HCl salt Form 2. In another embodiment,HCl Salt Form 2 has one, two, three, four, five, six, seven, eight,nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen,seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two,twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven,twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three,thirty-four, thirty-five, thirty-six, thirty-seven, or thirty-eightcharacteristic X-ray powder diffraction peaks as set forth in Table 35.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 69. In certainembodiments, the crystalline form, HCl salt Form 2, exhibits a TGAthermogram comprising a total mass loss of approximately 2.6% of thetotal mass of the sample between approximately 20° C. and approximately140° C. when heated from approximately 20° C. to approximately 325° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 69, where thecrystalline form HCl salt of Compound 1 exhibits a TGA thermogramcomprising a total mass loss of approximately 14% of the total mass ofthe sample between approximately 140° C. and approximately 200° C. whenheated from approximately 20° C. to approximately 325° C. Thus, incertain embodiments, the crystalline form HCl salt of Compound 1 losesabout 17% of its total mass when heated from about ambient temperatureto about 325° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram as depicted in FIG. 69 comprising anendothermic event with an onset temperature at about 151° C. and a peakmaximum temperature at about 157° C. when heated from approximately 25°C. to approximately 350° C.

In still another embodiment, HCl salt Form 2 is substantially pure. Incertain embodiments, the substantially pure HCl salt Form 2 issubstantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure HCl salt Form2 is no less than about 95%, no less than about 96%, no less than about97%, no less than about 98%, no less than about 98.5%, no less thanabout 99%, no less than about 99.5%, or no less than about 99.8%.

HCl Salt Form 3

In certain embodiments, provided herein is HCl salt Form 3.

In one embodiment, HCl salt Form 3 is a solid form of Compound 1. Inanother embodiment, HCl Salt Form 3 is crystalline. In one embodiment,HCl salt Form 3 is a solvated form of Compound 1. In one embodiment, HClsalt Form 3 is an n-butanol solvated form of Compound 1. In oneembodiment, HCl salt Form 3 is a heptane solvated form of Compound 1. Inone embodiment, HCl salt Form 3 is an n-butanol/heptane solvated form ofCompound 1. In one embodiment, HCl salt Form 3 is a hydrated form ofCompound 1. In one embodiment, HCl salt Form 3 is an anhydrate form ofCompound 1.

In certain embodiments, HCl salt Form 3 provided herein is obtained byequilibration experiments, evaporation experiments, coolingrecrystallization experiments and anti-solvent recrystallizationexperiments. In certain embodiments, HCl salt Form 3 is obtained fromcertain solvent systems including n-butanol, toluene, orn-butanol/toluene (about 1:1).

In certain embodiments, a solid form provided herein, e.g., HCl saltForm 3, is substantially crystalline, as indicated by, e.g., X-raypowder diffraction measurements. In one embodiment, the HCl salt Form 3has an X-ray powder diffraction pattern substantially as shown in FIG.70. In one embodiment, a solid form provided herein, e.g., HCl salt Form3, has one or more characteristic X-ray powder diffraction peaks atapproximately 5.5, 6.5, 7.7, 10.0, 10.5, 10.9, 12.9, 14.8, 15.9, 16.2,18.3, 18.9, 19.4, 20.4, 21.0, 21.8, 22.2, 22.5, 24.1, 26.0, 28.8, 29.9,32.7, or 39.4° 2θ (±0.2° 2θ) or (±0.1° 2θ) as depicted in FIG. 70. In aspecific embodiment, a solid form provided herein, e.g., HCl salt Form3, has one, two, three, four, five, six, seven, or eight characteristicX-ray powder diffraction peaks at approximately 5.5, 6.5, 10.9, 16.2,19.4, 20.4, 22.2, or 22.5° 2θ (±0.2° 2θ). In another embodiment, a solidform provided herein has one, two, three, or four characteristic X-raypowder diffraction peaks at approximately 5.5, 16.2, 19.4, or 20.4° 2θ(±0.2° 2θ). In one embodiment, the solid form is HCl Salt Form 3. Inanother embodiment, the HCl salt Form 3 has one, two, three, four, five,six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen,fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one,twenty-two, twenty-three, or twenty-four characteristic X-ray powderdiffraction peaks as set forth in Table 36.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 71. In certainembodiments, the crystalline form HCl salt of Compound 1, exhibits a TGAthermogram comprising a total mass loss of approximately 2.3% of thetotal mass of the sample between approximately 50° C. and approximately175° C. when heated from approximately 25° C. to approximately 140° C.In certain embodiments, the crystalline form HCl salt of Compound 1,exhibits a TGA thermogram comprising a total mass loss of approximately11% of the total mass of the sample between approximately 140° C. andapproximately 210° C. when heated from approximately 25° C. toapproximately 140° C. Thus, in certain embodiments, the crystalline formHCl salt of Compound 1 loses about 13% of its total mass when heatedfrom about ambient temperature to about 220° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram as depicted in FIG. 71 comprising anendothermic event with an onset temperature at about 153° C. and a peakmaximum temperature at about 168° C. when heated from approximately 25°C. to approximately 350° C.

In still another embodiment, HCl salt Form 3 is substantially pure. Incertain embodiments, the substantially pure HCl salt Form 3 issubstantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure HCl salt Form3 is no less than about 95%, no less than about 96%, no less than about97%, no less than about 98%, no less than about 98.5%, no less thanabout 99%, no less than about 99.5%, or no less than about 99.8%.

HCl Salt Form 4

In certain embodiments, provided herein is HCl salt Form 4.

In one embodiment, HCl salt Form 4 is a solid form of Compound 1. Inanother embodiment, HCl salt Form 4 is crystalline. In one embodiment,HCl salt Form 4 is a solvated form of Compound 1. In one embodiment, HClsalt Form 4 is a methanol solvated form of Compound 1. In oneembodiment, HCl salt Form 4 is an IPAc solvated form of Compound 1. Inone embodiment, HCl salt Form 4 is a MeOH/IPAc solvated form of Compound1.

In certain embodiments, HCl salt Form 4 provided herein is obtained byequilibration experiments, evaporation experiments, coolingrecrystallization experiments and anti-solvent recrystallizationexperiments. In certain embodiments, HCl salt Form 4 is obtained fromcertain solvent systems including MeOH/IPAc.

In certain embodiments, a solid form provided herein, e.g., HCl saltForm 4, is substantially crystalline, as indicated by, e.g., X-raypowder diffraction measurements. In one embodiment, a solid formprovided herein, e.g., HCl salt Form 4, has an X-ray powder diffractionpattern substantially as shown in FIG. 72. In one embodiment, a solidform provided herein, e.g., HCl salt Form 4, has one or morecharacteristic X-ray powder diffraction peaks at approximately 7.9, 8.1,8.2, 8.4, 10.8, 14.0, 15.3, 15.9, 16.4, 16.8, 17.8, 18.3, 18.9, 19.1,19.7, 20.3, 21.0, 21.6, 22.1, 23.3, 23.6, 24.9, 25.3, 26.3, 27.6, 28.5,29.1, 29.9, 30.6, 30.9, 31.9, 32.8, 34.6, or 36.2° 2θ (±0.2° 2θ) or(±0.1° 2θ) as depicted in FIG. 72. In a specific embodiment, a solidform provided herein, e.g., HCl salt Form 4, has one, two, three, four,five, six, seven or eight characteristic X-ray powder diffraction peaksat approximately 7.9, 8.1, 8.4, 15.9, 16.8, 18.9, 19.1, or 19.7° 2θ(±0.2° 2θ). In another embodiment, a solid form provided herein has one,two, three, or four characteristic X-ray powder diffraction peaks atapproximately 7.9, 8.1, 8.4, or 15.9° 2θ (±0.2° 2θ). In one embodiment,the solid form is HCl Salt Form 4. In another embodiment, HCl salt Form4 has one, two, three, four, five, six, seven, eight, nine, ten, eleven,twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen,nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four,twenty-five, twenty-six, twenty-seven, twenty-eight, twenty-nine,thirty, thirty-one, thirty-two, thirty-three, or thirty-fourcharacteristic X-ray powder diffraction peaks as set forth in Table 37.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 73. In certainembodiments, the crystalline form HCl salt of Compound 1 exhibits a TGAthermogram comprising a total mass loss of approximately 4.5% of thetotal mass of the sample between approximately 20° C. and approximately140° C. when heated from approximately 20° C. to approximately 275° C.Thus, in certain embodiments, the crystalline form HCl salt of Compound1 loses about 4.5% of its total mass when heated from about ambienttemperature to about 275° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram as depicted in FIG. 73 comprising anendothermic event with an onset temperature at about 94° C. and a peakmaximum temperature at about 118° C. when heated from approximately 25°C. to approximately 275° C. In one embodiment, provided herein is acrystalline form HCl salt of Compound 1 having a DSC thermogram asdepicted in FIG. 73 further comprising an endothermic event with anonset temperature at about 219° C. and a peak maximum temperature atabout 236° C. when heated from approximately 25° C. to approximately275° C.

In still another embodiment, HCl salt Form 4 is substantially pure. Incertain embodiments, the substantially pure HCl salt Form 4 issubstantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure HCl salt Form4 is no less than about 95%, no less than about 96%, no less than about97%, no less than about 98%, no less than about 98.5%, no less thanabout 99%, no less than about 99.5%, or no less than about 99.8%.

HCl Salt Form 5

In certain embodiments, provided herein is HCl salt Form 5.

In one embodiment, HCl salt Form 5 is a solid form of Compound 1. Inanother embodiment, HCl salt Form 5 is crystalline. In one embodiment,HCl salt Form 5 is a solvated form of Compound 1. In one embodiment, HClsalt Form 5 is a DMF solvated form of Compound 1.

In certain embodiments, HCl salt Form 5 provided herein is obtained byequilibration experiments, vapor diffusion, and evaporation experiments.In certain embodiments, HCl salt Form 5 is obtained from certain solventsystems including DMF.

In certain embodiments, a solid form provided herein, e.g., HCl saltForm 5, is substantially crystalline, as indicated by, e.g., X-raypowder diffraction measurements. In one embodiment, a solid formprovided herein, e.g., HCl salt Form 5, has an X-ray powder diffractionpattern substantially as shown in FIG. 74. In one embodiment, a solidform provided herein, e.g., HCl salt Form 5, has one or morecharacteristic X-ray powder diffraction peaks at approximately 7.9, 8.7,10.0, 11.7, 13.3, 13.6, 15.1, 15.7, 17.2, 17.9, 19.9, 20.6, 21.3, 23.3,24.2, 25.5, 27.0, 28.5, 29.3, 30.1, 31.7, 32.2, 34.1, 35.4, 37.0, or38.8° 2θ (±0.2° 2θ) or (±0.1° 2θ) as depicted in FIG. 74. In a specificembodiment, a solid form provided herein, e.g., HCl salt Form 5, hasone, two, three, four, five, six, seven, eight, nine, ten, eleven, ortwelve characteristic X-ray powder diffraction peaks at approximately7.9, 8.7, 10.0, 11.7, 15.1, 15.7, 17.2, 19.9, 20.6, 21.3, 24.2, or 27.0°2θ (±0.2° 2θ). In another embodiment, a solid form provided herein hasone, two, three, or four characteristic X-ray powder diffraction peaksat approximately 7.9, 19.9, 20.6, or 27.0° 2θ (±0.2° 2θ). In oneembodiment, the solid form is HCl Salt Form 5. In another embodiment,HCl salt Form 5 has one, two, three, four, five, six, seven, eight,nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen,seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two,twenty-three, twenty-four, twenty-five, or twenty-six characteristicX-ray powder diffraction peaks as set forth in Table 38.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 75. In certainembodiments, the crystalline form HCl salt of Compound 1 exhibits a TGAthermogram comprising a total mass loss of approximately 4.2% of thetotal mass of the sample between approximately 25° C. and approximately140° C. when heated from approximately 25° C. to approximately 300° C.Thus, in certain embodiments, the crystalline form HCl salt of Compound1 loses about 4.2% of its total mass when heated from about ambienttemperature to about 300° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 75 comprising an endothermicevent with a maximum at about 104° C. when heated from approximately 25°C. to approximately 220° C.

In one embodiment, provided herein is a crystalline form of Compound 1having a DSC thermogram as depicted in FIG. 75 comprising an endothermicevent with an onset temperature at about 192° C. and a peak maximumtemperature at about 209° C. when heated from approximately 25° C. toapproximately 220° C.

In still another embodiment, HCl salt Form 5 is substantially pure. Incertain embodiments, the substantially pure HCl salt Form 5 issubstantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure HCl salt Form5 is no less than about 95%, no less than about 96%, no less than about97%, no less than about 98%, no less than about 98.5%, no less thanabout 99%, no less than about 99.5%, or no less than about 99.8%.

HCl Salt Form 6

In certain embodiments, provided herein is HCl salt Form 6.

In one embodiment, HCl salt Form 6 is a solid form of Compound 1. Inanother embodiment, HCl salt Form 6 is crystalline. In one embodiment,HCl salt Form 6 is a hydrate of Compound 1. In one embodiment, HCl saltForm 6 is a pentahydrate of Compound 1.

In certain embodiments, HCl salt Form 6 provided herein is obtained byequilibration experiments. In certain embodiments, HCl salt Form 6 isobtained from certain solvent systems including 0.1N HCl in water.

In certain embodiments, a solid form provided herein, e.g., the HCl saltForm 6, is substantially crystalline, as indicated by, e.g., X-raypowder diffraction measurements. In one embodiment, a solid formprovided herein, e.g., the HCl salt Form 6, has an X-ray powderdiffraction pattern substantially as shown in FIG. 76. In oneembodiment, a solid form provided herein, e.g., the HCl salt Form 6, hasone or more characteristic X-ray powder diffraction peaks atapproximately 7.0, 8.8, 9.9, 11.5, 12.2, 12.4, 14.7, 16.3, 16.7, 17.5,17.9, 18.2, 18.6, 19.2, 19.4, 19.7, 19.9, 20.3, 21.0, 21.2, 21.9, 22.7,22.9, 23.7, 24.6, 25.0, 25.6, 26.3, 26.5, 26.8, 27.2, 27.6, 28.2, 28.7,29.1, 29.6, 30.3, 30.8, 31.2, 31.7, 32.3, 32.8, 33.3, 34.0, 34.3, 35.3,36.2, or 37.0° 2θ (±0.2° 2θ) or (±0.1° 2θ) as depicted in FIG. 76. In aspecific embodiment, a solid form provided herein, e.g., the HCl saltForm 6, has one, two, three, four, five, six, seven, eight, nine, ten,eleven, or twelve characteristic X-ray powder diffraction peaks atapproximately 9.9, 12.4, 16.3, 17.5, 19.2, 19.7, 19.9, 21.0, 25.0, 25.6,27.2, or 32.3° 2θ (±0.2° 2θ). In another embodiment, a solid formprovided herein has one, two, three, or four characteristic X-ray powderdiffraction peaks at approximately 9.9, 12.4, 17.9, or 19.7° 2θ (±0.2°2θ). In one embodiment, the solid form is HCl Salt Form 6. In anotherembodiment, HCl salt Form 6 has one, two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen,seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two,twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven,twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three,thirty-four, thirty-five, thirty-six, thirty-seven, thirty-eight,thirty-nine, forty, forty-one, forty-two, forty-three, forty-four,forty-five, forty-six, forty-seven, or forty-eight characteristic X-raypowder diffraction peaks as set forth in Table 39.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 77. In certainembodiments, the crystalline form HCl salt of Compound 1 exhibits a TGAthermogram comprising a total mass loss of approximately 11.6% of thetotal mass of the sample between approximately 25° C. and approximately100° C. when heated from approximately 20° C. to approximately 300° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 78. In certainembodiments, the crystalline form HCl salt of Compound 1 exhibits a TGAthermogram comprising a total mass loss of approximately 14.3% of thetotal mass of the sample between approximately 25° C. and approximately60° C. when heated from approximately 20° C. to approximately 300° C.

In certain embodiments, the crystalline form HCl salt of Compound 1exhibits a TGA thermogram comprising a total mass loss of approximately1.6% of the total mass of the sample between approximately 60° C. andapproximately 125° C. when heated from approximately 20° C. toapproximately 300° C. Thus, in certain embodiments, the crystalcrystalline form HCl salt of Compound 1 loses about 15.9% of its totalmass when heated from about ambient temperature to about 100° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram as depicted in FIG. 77 comprising anendothermic event with an onset temperature at about 53° C. and a peakmaximum temperature at about 88° C. when heated from approximately 25°C. to approximately 325° C. In one embodiment, provided herein is acrystalline form HCl salt of Compound 1 having a DSC thermogram asdepicted in FIG. 77 comprising an endothermic event with an onsettemperature at about 201° C. and a peak maximum temperature at about228° C. when heated from approximately 25° C. to approximately 325° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram as depicted in FIG. 79 comprising anendothermic event with an onset temperature at about 59° C. and a peakmaximum temperature at about 89° C. when heated from approximately 25°C. to approximately 325° C. In one embodiment, provided herein is acrystalline form HCl salt of Compound 1 having a DSC thermogram asdepicted in FIG. 79 comprising an endothermic event with an onsettemperature at about 211° C. and a peak maximum temperature at about225° C. when heated from approximately 25° C. to approximately 325° C.

In still another embodiment, HCl salt Form 6 is substantially pure. Incertain embodiments, the substantially pure HCl salt Form 6 issubstantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure HCl salt Form6 is no less than about 95%, no less than about 96%, no less than about97%, no less than about 98%, no less than about 98.5%, no less thanabout 99%, no less than about 99.5%, or no less than about 99.8%.

HCl Salt Form 7

In certain embodiments, provided herein is HCl salt Form 7.

In one embodiment, HCl salt Form 7 is a solid form of Compound 1. Inanother embodiment, HCl salt Form 7 is crystalline. In one embodiment,HCl salt Form 7 is a hydrate of Compound 1. In one embodiment, HCl saltForm 7 is a monohydrate of Compound 1.

In certain embodiments, HCl salt Form 7 provided herein is obtained byequilibration experiments, evaporation experiments and anti-solventrecrystallization experiments. In certain embodiments, HCl salt Form 7is obtained from certain solvent systems including water at roomtemperature.

In certain embodiments, a solid form provided herein, e.g., HCl saltForm 7, is substantially crystalline, as indicated by, e.g., X-raypowder diffraction measurements. In one embodiment, a solid formprovided herein, e.g., HCl salt Form 7, has an X-ray powder diffractionpattern substantially as shown in FIG. 80. In one embodiment, a solidform provided herein, e.g., HCl salt Form 7, has one or morecharacteristic X-ray powder diffraction peaks at approximately 7.9, 8.1,8.3, 10.8, 13.8, 14.5, 15.3, 15.6, 16.2, 16.6, 17.0, 17.6, 18.3, 18.6,19.1, 19.6, 19.7, 20.2, 20.7, 21.5, 22.0, 22.9, 24.0, 24.3, 25.2, 26.2,28.4, 29.0, 29.5, 30.2, 30.8, 31.2, 32.5, 33.1, 34.7, or 36.3° 2θ (±0.2°2θ) or (±0.1° 2θ) as depicted in FIG. 80. In a specific embodiment, asolid form provided herein, e.g., HCl salt Form 7, has one, two, three,four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, orfourteen characteristic X-ray powder diffraction peaks at approximately7.9, 8.1, 8.3, 10.8, 15.3, 16.2, 18.3, 19.1, 19.6, 19.7, 21.5, 24.0,24.3, or 25.2° 2θ (±0.2° 2θ). In another embodiment, a solid formprovided herein has one, two, three, or four characteristic X-ray powderdiffraction peaks at approximately 8.1, 8.3, 19.1, or 19.7° 2θ (±0.2°2θ). In one embodiment, the solid form is HCl Salt Form 7. In anotherembodiment, HCl salt Form 7 has one, two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen,seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two,twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven,twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three,thirty-four, thirty-five, or thirty-six characteristic X-ray powderdiffraction peaks as set forth in Table 40.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 81. In certainembodiments, the crystalline form HCl salt of Compound 1 exhibits a TGAthermogram comprising a total mass loss of approximately 3.8% of thetotal mass of the sample between approximately 25° C. and approximately100° C. when heated from approximately 25° C. to approximately 300° C.Thus, in certain embodiments, the crystalline form HCl salt of Compound1 loses about 3.8% of its total mass when heated from about ambienttemperature to about 300° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 83. In certainembodiments, the crystalline form HCl salt of Compound 1 exhibits a TGAthermogram comprising a total mass loss of approximately 3.4% of thetotal mass of the sample between approximately 25° C. and approximately100° C. when heated from approximately 25° C. to approximately 300° C.Thus, in certain embodiments, the crystalline form HCl salt of Compound1 loses about 3.4% of its total mass when heated from about ambienttemperature to about 300° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram as depicted in FIG. 81 comprising anendothermic event with an onset temperature at about 80° C. and a peakmaximum temperature at about 111° C. when heated from approximately 25°C. to approximately 300° C. In one embodiment, provided herein is acrystalline form HCl salt of Compound 1 having a DSC thermogram asdepicted in FIG. 81 comprising an endothermic event with an onsettemperature at about 215° C. and a peak maximum temperature at about233° C. when heated from approximately 25° C. to approximately 300° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram as depicted in FIG. 82 comprising anendothermic event with an onset temperature at about 71° C. and a peakmaximum temperature at about 98° C. when heated from approximately 25°C. to approximately 300° C. In one embodiment, provided herein is acrystalline form HCl salt of Compound 1 having a DSC thermogram asdepicted in FIG. 82 comprising an endothermic event with an onsettemperature at about 209° C. and a peak maximum temperature at about230° C. when heated from approximately 25° C. to approximately 300° C.

In one embodiment, provided herein is a solid form, e.g., a startingmaterial HCl salt Form, having a DVS isotherm plot substantially asdepicted in FIG. 84.

In still another embodiment, HCl salt Form 7 is substantially pure. Incertain embodiments, the substantially pure HCl salt Form 7 issubstantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure HCl salt Form7 is no less than about 95%, no less than about 96%, no less than about97%, no less than about 98%, no less than about 98.5%, no less thanabout 99%, no less than about 99.5%, or no less than about 99.8%.

HCl Salt Form 8

In certain embodiments, provided herein is HCl salt Form 8.

In one embodiment, HCl salt Form 8 is a solid form of Compound 1. Inanother embodiment, HCl salt Form 8 is crystalline. In one embodiment,HCl salt Form 8 is a hydrate of Compound 1. In one embodiment, HCl saltForm 8 is a monohydrate of Compound 1.

In certain embodiments, HCl salt Form 8 provided herein is obtained byequilibration experiments, evaporation experiments and anti-solventrecrystallization experiments. In certain embodiments, HCl salt Form 8is obtained from certain solvent systems including water at 50° C.

In certain embodiments, a solid form provided herein, e.g., HCl saltForm 8, is substantially crystalline, as indicated by, e.g., X-raypowder diffraction measurements. In one embodiment, a solid formprovided herein, e.g., HCl salt Form 8, has an X-ray powder diffractionpattern substantially as shown in FIG. 85. In one embodiment, a solidform provided herein, e.g., HCl salt Form 8 has one or morecharacteristic X-ray powder diffraction peaks at approximately 8.1, 9.8,10.1, 10.8, 11.1, 11.6, 15.7, 16.2, 16.9, 17.4, 18.0, 18.7, 19.2, 19.6,21.4, 22.1, 22.9, 23.8, 24.2, 25.1, 25.5, 26.2, 26.7, 28.2, 28.4, 29.6,30.5, 31.7, 32.0, 33.7, 35.6, 36.4, or 37.4° 2θ (±0.2° 2θ) or (±0.1° 2θ)as depicted in FIG. 85. In a specific embodiment, a solid form providedherein has one, two, three, or four characteristic X-ray powderdiffraction peaks at approximately 9.8, 17.4, 18.7, or 26.2° 2θ (±0.2°2θ). In one embodiment, the solid form is HCl Salt Form 8. In anotherembodiment, HCl salt Form 8 has one, two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen,seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two,twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven,twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, orthirty-three characteristic X-ray powder diffraction peaks as set forthin Table 41.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 86. In certainembodiments, the crystalline form HCl salt of Compound 1 exhibits a TGAthermogram comprising a total mass loss of approximately 3.1% of thetotal mass of the sample between approximately 25° C. and approximately100° C. when heated from approximately 25° C. to approximately 300° C.In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a TGA thermograph corresponding substantially to therepresentative TGA thermogram as depicted in FIG. 87. In certainembodiments, the crystalline form HCl salt of Compound 1 exhibits a TGAthermogram comprising a total mass loss of approximately 3.0% of thetotal mass of the sample between approximately 30° C. and approximately120° C. when heated from approximately 25° C. to approximately 300° C.

In certain embodiments, the crystalline form HCl salt of Compound 1exhibits a TGA thermogram comprising a total mass loss of approximately2.6% of the total mass of the sample between approximately 125° C. andapproximately 215° C. when heated from approximately 30° C. toapproximately 300° C. Thus, in certain embodiments, the crystalline formHCl salt of Compound 1 loses about 5.6% of its total mass when heatedfrom about ambient temperature to about 220° C. The theoretical watercontent for the monohydrate HCl salt Form 8 is 2.9% and matches thepercent total mass lost by the sample in the above TGA thermogram.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram as depicted in FIG. 86 comprising anendothermic event with an onset temperature at about 117° C. and a peakmaximum temperature at about 148° C. when heated from approximately 25°C. to approximately 300° C. In one embodiment, provided herein is acrystalline form HCl salt of Compound 1 having a DSC thermogram asdepicted in FIG. 86 comprising an endothermic event with an onsettemperature at about 208° C. and a peak maximum temperature at about221° C. when heated from approximately 25° C. to approximately 300° C.

In one embodiment, provided herein is a crystalline form HCl salt ofCompound 1 having a DSC thermogram as depicted in FIG. 88 comprising anendothermic event with a maximum at about 148° C. when heated fromapproximately 25° C. to approximately 275° C. In one embodiment,provided herein is a crystalline form HCl salt of Compound 1 having aDSC thermogram as depicted in FIG. 88 comprising an endothermic eventwith an onset temperature at about 204° C. and a peak maximumtemperature at about 224° C. when heated from approximately 25° C. toapproximately 300° C.

In still another embodiment, HCl salt Form 8 is substantially pure. Incertain embodiments, the substantially pure HCl salt Form 8 issubstantially free of other solid forms, e.g., amorphous solid. Incertain embodiments, the purity of the substantially pure HCl salt Form8 is no less than about 95%, no less than about 96%, no less than about97%, no less than about 98%, no less than about 98.5%, no less thanabout 99%, no less than about 99.5%, or no less than about 99.8%.

Methods of Use

The solid forms of Compound 1 described herein have utility aspharmaceuticals to treat, prevent or improve conditions in animals orhumans. Accordingly, provided herein are solid forms of Compound 1described herein that can be used in all the methods as provided herein.Particularly, the solid forms of Compound 1 as provided herein are foruses in the treatment or prevention of a cancer. The methods providedherein comprise the administration of an effective amount of one or moresolid forms of Compound 1 described herein to a subject in need thereof.It is to be understood that the methods described herein also includetreatment with a pharmaceutical composition, such as those providedbelow, where the pharmaceutical composition includes a solid form ofCompound 1 described herein and optionally at least one pharmaceuticallyacceptable excipient.

In another aspect, provided herein are methods for treating orpreventing a cancer, comprising administering to a subject in needthereof an effective amount of a solid form of Compound 1 a solid formof Compound 1, as described herein. In some embodiments, the cancer is asolid tumor or a hematological tumor. In some embodiments, the cancer isnot melanoma.

In some embodiments, the solid tumor is melanoma, colorectal cancer,stomach cancer, head and neck cancer, thyroid cancer, bladder cancer,CNS cancer, lung cancer, pancreatic cancer, and soft tissue cancer. Inone embodiment, the solid tumor is endocrine cancer, bladder cancer,breast cancer, cervix cancer, colon cancer, duodenum cancer, glioma,head and d neck cancer, kidney cancer, liver cancer, lung cancer (e.g.non-small cell lung cancer NSCLC), esophageal cancer, thyroid cancer, orpancreatic cancer.

In other embodiment, the cancer is bladder cancer, breast cancer (forexample Her positive, Her negative, or EGFR positive), CNS cancer(including neuroblastoma, and glioma), colon cancer, gastrointestinalcancer (for example, stomach cancer, and colon cancer), endocrine cancer(for example, thyroid cancer, or adrenal gland cancer), femalegenitoureal cancer (for example, cervix cancer, ovary clear cell cancer,vulva cancer, uterus cancer, or ovary cancer), head and neck cancer,hematopoietic cancer (for example, leukemia or myeloma), kidney cancer,liver cancer, lung cancer (for example, NSCLC, or SCLC), melanoma,pancreas cancer, prostate cancer, or soft tissue cancer (for example,sarcoma, or osteosarcoma).

In another embodiment, the cancer is bladder cancer, breast cancer (forexample Her positive, Her negative, or EGFR positive), CNS cancer (forexample, glioma, or neuroblastoma), colon cancer, gastrointestinalcancer (for example, stomach cancer), endocrine cancer (for example,thyroid cancer or adrenal gland cancer), female genitoureal cancer (forexample, cancer of the uterus, cervix, ovary clear cell, or vulva), headand neck cancer, hematopoietic cancer (for example, leukemia ormyeloma), kidney cancer, liver cancer, lung cancer (for example, NSCLC,or SCLC), melanoma, pancreas cancer, prostate cancer, or soft tissuecancer (for example, sarcoma or osteosarcoma).

In still another embodiment, the cancer is a cancer set forth in Table44.

Also provided herein are methods for treating or preventinghepatocellular carcinoma (HCC), comprising administering to a subject inneed thereof an effective amount of a solid form of Compound 1, asdescribed herein.

Also provided herein are methods for treating or preventing colorectalcancer (CRC), melanoma, gastric cancer, HCC, lung cancer, pancreaticcancer, leukemia, or multiple myeloma, comprising administering to asubject in need thereof an effective amount of a solid form of Compound1 as described herein or a pharmaceutical composition thereof, asdescribed herein. In one embodiment, the CRC, gastric, or HCC is acancer characterized by a β-catenin mutation. Also provided herein aremethods for treating or preventing colorectal cancer (CRC), gastriccancer, HCC, lung cancer, pancreatic cancer, leukemia, and multiplemyeloma, comprising administering to a subject in need thereof aneffective amount of a solid form of Compound 1 as described herein, asdescribed herein.

In another embodiment provided herein are methods of treating leukemiacomprising administering a solid form of Compound 1 as described hereinor a pharmaceutical composition thereof. The leukemia can be chronicmyelogenous leukemia (CIVIL). In another embodiment, the leukemia isacute myelogenous leukemia (AML). In one embodiment, the leukemia isFLT-3 mutated AML.

In another embodiment provided herein are methods of treating lymphomacomprising administering a solid form of Compound 1 as described hereinor a pharmaceutical composition thereof. The lymphoma can be Burkitt'slymphoma. In one embodiment, the leukemia is Hodgkin's lymphoma. Inanother embodiment, the leukemia is a B-cell lymphoma. In anotherembodiment, the leukemia is a T-cell lymphoma. In still anotherembodiment, the lymphoma is primary effusion lymphoma (PEL).

The solid forms of Compound 1) show anti-proliferative activity in avariety of cancer cell lines. (Table 44) Anti-proliferative activity inthese cancer cell lines indicates that the solid forms of Compound 1 areuseful in the treatment of cancers, including hematopoietic and solidtumors. In one embodiment, the hematopoietic and solid tumors areselected from bladder cancer, breast cancer, CNS cancer (for example,neuroblastoma, medulloblastoma and glioma), colon cancer, duodenumcancer, endocrine cancer (for example, thyroid cancer and adrenal glandcancer), female genitourinary cancer (for example, uterus cancer, cervixcancer, ovary cancer and vulva cancer), head and neck cancer (forexample, esophageal cancer), hematopoietic and lymphoid cancer (forexample, lymphoma, leukemia, and myeloma), kidney cancer, liver cancer,lung cancer (for example, NSCLC and SCLC), pancreas cancer, prostatecancer, skin cancer (for example, melanoma and carcinoma), soft tissuecancer (for example, sarcoma and osteosarcoma), stomach cancer, andtestis cancer. In one embodiment, the hematopoietic and solid tumors areselected from bladder cancer, breast cancer, CNS cancer (for example,neuroblastoma, medulloblastoma and glioma), colon cancer, duodenumcancer, endocrine cancer (for example, thyroid cancer and adrenal glandcancer), female genitourinary cancer (for example, uterus cancer, cervixcancer, and vulva cancer), head and neck cancer, hematopoietic andlymphoid cancer (for example, lymphoma, leukemia, and myeloma), kidneycancer, liver cancer, lung cancer (for example, NSCLC and SCLC),pancreas cancer, prostate cancer, skin cancer (for example, melanoma andcarcinoma), soft tissue cancer (for example, sarcoma and osteosarcoma),stomach cancer, and testis cancer.

In another embodiment, the solid forms of Compound 1 described hereininduce apoptosis in a variety of cancer cell lines. Induction ofapoptosis indicates that the solid forms of Compound 1 described hereinare useful in the treatment of cancers, including hematopoietic andsolid tumors. In one embodiment, the hematopoietic and solid tumors areselected from bladder cancer, breast cancer, CNS cancer (for example,neuroblastoma, and glioma), colon cancer, duodenum cancer, endocrinecancer (for example, thyroid cancer and adrenal gland cancer), femalegenitourinary cancer (for example, uterus cancer, cervix cancer, ovarycancer and vulva cancer), head and neck cancer (for example, esophagealcancer), hematopoietic and lymphoid cancer (for example, lymphoma,leukemia, and myeloma), kidney cancer, liver cancer, lung cancer (forexample, NSCLC and SCLC), pancreas cancer, prostate cancer, skin cancer(for example, melanoma and carcinoma), soft tissue cancer (for example,sarcoma and osteosarcoma), stomach cancer, and testis cancer. In oneembodiment, the hematopoietic and solid tumors are selected from bladdercancer, breast cancer, CNS cancer (for example, neuroblastoma, andglioma), colon cancer, duodenum cancer, endocrine cancer (for example,thyroid cancer and adrenal gland cancer), female genitourinary cancer(for example, vulva cancer), head and neck cancer (for example,esophageal cancer), hematopoietic and lymphoid cancer (for example,lymphoma, and leukemia), kidney cancer, liver cancer, lung cancer (forexample, NSCLC and SCLC), pancreas cancer, prostate cancer, skin cancer(for example, melanoma), soft tissue cancer (for example, sarcoma andosteosarcoma), stomach cancer, and testis cancer. In one embodiment, thehematopoietic and solid tumors are selected from bladder cancer, breastcancer, CNS cancer (for example, medulloblastoma, neuroblastoma, andglioma), colon cancer, duodenum cancer, endocrine cancer (for example,thyroid cancer and adrenal gland cancer), female genitourinary cancer(for example, placenta cancer, uterus cancer, cervix cancer, ovarycancer and vulva cancer), head and neck cancer (for example, esophagealcancer), hematopoietic and lymphoid cancer (for example, lymphoma,leukemia, and myeloma), kidney cancer, liver cancer, lung cancer (forexample, NSCLC and SCLC), pancreas cancer, prostate cancer, skin cancer(for example, melanoma and carcinoma), soft tissue cancer (for example,sarcoma and osteosarcoma), stomach cancer, and testis cancer. In stillanother embodiment, the cases is a cancer set forth in Table 44.

Also provided herein are methods for treating or preventing a cancercharacterized by a BRAF mutation and/or a beta-catenin mutation(alternatively referred to as CTNNB1 mutation), comprising administeringto a subject in need thereof an effective amount of a solid form ofCompound 1, as described herein. In some such embodiments, the cancer ischaracterized by a BRAF mutation. In another embodiment, the cancer ischaracterized by a beta-catenin mutation. In yet another embodiment, thecancer is characterized by an activated beta-catenin pathway. In somesuch embodiments, the cancer is CRC or melanoma characterized by a BRAFmutation. In other embodiments, the cancer is CRC characterized by abeta-catenin mutation, additionally comprising an EGFR mutation orincreased EGFR activity (for example, CRC characterized by an activatedbeta-catenin pathway and an EGFR mutation, or CRC characterized by anactivated beta-catenin pathway and increased EGFR activity). In stillother embodiments, the cancer is gastric cancer characterized by abeta-catenin mutation, additionally comprising a KRAS mutation (i.e.gastric cancer characterized by an activated beta-catenin pathway and aKRAS mutation). In another embodiment the cancer is HCC, characterizedby an activated beta-catenin pathway. In some such embodiments, the BRAFmutation is BRAF V660E. In some such embodiments, the BRAF mutation isBRAF V600E. In some such embodiments, the BRAF mutation is one or moreof BRAF V600E, BRAF T119S, or BRAF G596R. In some such embodiments, thebeta-catenin mutation is one or more of beta-catenin S33Y, G34E, S45del,or S33C. In some such embodiments, the EGFR mutation is one or more ofEGFR E282K, G719S, P753S, or V1011M. In some such embodiments, the KRASmutation is A146T, G12C, G12D, G12V, G13D, or Q61L.

Also provided herein are methods for treating or preventing a cancerexpressing PD-L1, comprising administering to a subject in need thereofan effective amount of a solid form of Compound 1, as described herein.In some such embodiments, the PD-L1 expressing cancer is melanoma, lungcancer, renal cell carcinoma (RCC), or HCC.

Also provided herein are methods for treating or preventing a cancercharacterized by a BRAF mutation, comprising administering to a subjectin need thereof an effective amount of a solid form of Compound 1, asdescribed herein. In some such embodiments, the cancer characterized bya BRAF mutation is CRC, thyroid cancer, melanoma or lung cancer. In somesuch embodiments, the cancer characterized by a BRAF mutation is CRC,thyroid cancer, or lung cancer. In some such embodiments, the BRAFmutation is BRAF V660E. In some such embodiments, the BRAF mutation isBRAF V600E. In other embodiments, the BRAF mutation is one or more ofBRAF V600E, BRAF T119S, or BRAF G596R.

Also provided herein are methods for treating or preventing a cancercharacterized by an NRAS mutation, comprising administering to a subjectin need thereof an effective amount of a solid form of Compound 1, asdescribed herein. In some such embodiments, the cancer characterized byan NRAS mutation is melanoma.

Also provided herein are methods for treating or preventing a cancercharacterized by a KRAS mutation, comprising administering to a subjectin need thereof an effective amount of a solid form of Compound 1, asdescribed herein. In some such embodiments, the cancer characterized bya KRAS mutation is CRC, pancreas cancer or lung cancer.

Also provided herein are methods for treating or preventing a cancercharacterized by a beta-catenin mutation, comprising administering to asubject in need thereof an effective amount of a solid form of Compound1, as described herein. Also provided herein are methods for treating orpreventing a cancer characterized by an activated beta-catenin pathway,comprising administering to a subject in need thereof an effectiveamount of a solid form of Compound 1, as described herein. In some suchembodiments, the cancer characterized by a beta-catenin mutation is CRC,stomach cancer, HCC or sarcoma. In some such embodiments, the cancercharacterized by an activated beta-catenin pathway is CRC, stomachcancer, HCC or sarcoma.

Also provided herein are methods for treating or preventinghepatocellular carcinoma (HCC), comprising administering to a subject inneed thereof an effective amount of a solid form of Compound 1, asdescribed herein. In some such embodiments, the HCC is characterized bya beta-catenin mutation and/or increased YAP expression. In some suchembodiments, the HCC is characterized by an activated beta-cateninpathway and/or increased YAP amplification expression. In someembodiments, the increased YAP expression is due to amplification or amutation.

Also provided herein are methods for treating or preventing colorectalcancer (CRC), comprising administering to a subject in need thereof aneffective amount of a solid form of Compound 1, as described herein. Insome such embodiments, the CRC is characterized by a BRAF mutationand/or beta-catenin mutation. In some such embodiments, the CRC ischaracterized by a BRAF mutation and/or an activated beta-cateninpathway.

Also provided herein are methods for treating or preventing gastriccancer, comprising administering to a subject in need thereof aneffective amount of a solid form of Compound 1, as described herein. Insome such embodiments, the gastric cancer is characterized by abeta-catenin mutation. In some such embodiments, the gastric cancer ischaracterized by an activated beta-catenin pathway.

Also provided herein are methods for treating or preventing melanoma,comprising administering to a subject in need thereof an effectiveamount of a solid form of Compound 1, as described herein. In some suchembodiments, the melanoma is characterized by a BRAF mutation and/orNRAS mutation.

Further provided herein are methods for predicting response to treatmentwith a solid form of Compound 1 described herein in a patient having acancer characterized by a gene mutation, the method comprising: a)obtaining a biological test sample from the patient's cancer; b)obtaining the gene sequence of one or more genes selected from BRAF,NRAS, KRAS, and/or CTNNB1 in said biological test sample; c) comparingsaid gene sequence(s) to the gene sequence(s) of a biological wild-typesample; wherein the presence of a mutation indicates an increasedlikelihood of response to a solid form of Compound 1 described hereintreatment of said patient's cancer. In some such embodiments, the methodadditionally comprises administering an effective amount of a solid formof Compound 1, as described herein.

Further provided herein are methods for predicting therapeutic efficacyof a solid form of Compound 1 described herein for treatment of apatient having a cancer characterized by a gene mutation, the methodcomprising: a) obtaining a biological test sample from the patient'scancer; b) obtaining the gene sequence(s) of one or more genes selectedfrom BRAF, NAS, KRAS, and/or CTNNB1 in said biological test sample; c)comparing said gene sequence(s) to the gene sequence(s) of a biologicalwild-type sample; wherein the presence of a mutation indicates anincreased likelihood of therapeutic efficacy of said treatment with asolid form of Compound 1 described herein for said patient. In some suchembodiments, the method additionally comprises administering aneffective amount of a solid form of Compound 1, as described herein.

In some embodiments, provided herein are methods for treating andpreventing cancer metastasis, comprising administering to a subject inneed thereof an effective amount of a solid form of Compound 1, asdescribed herein. In some embodiments, the cancer is a metastaticcancer, in particular, a metastatic solid tumor or metastatichematologic cancer, wherein the solid tumor and hematologic cancer is asdescribed herein. In other embodiments, provided herein are methods oftreating and preventing cancer metastasis, comprising administering to asubject in need thereof an effective amount of a solid form of Compound1, as described herein. In yet another aspect, provided herein ismethods of eradicating cancer stem cells in a subject, comprisingadministering to a subject in need thereof an effective amount of asolid form of Compound 1, as described herein. In other embodiments,provided herein are methods of inducing differentiation in cancer stemcells in a subject, comprising administering to a subject in needthereof an effective amount of a solid form of Compound 1, as describedherein. In other embodiments, provided herein are methods of inducingcancer stem cell death in a subject, comprising administering to asubject in need thereof an effective amount of a solid form of Compound1, as described herein. In some such embodiments, the cancer is a solidtumor or a hematological cancer, as described herein.

In one embodiment, provided herein are methods for achieving a ResponseEvaluation Criteria in Solid Tumors (RECIST 1.1) of complete response,partial response or stable disease in a patient comprising administeringan effective amount of a solid form of Compound 1 described herein to apatient having a cancer, in particular a solid tumor as describedherein. In another embodiment, provided herein are methods to increaseProgression Free Survival rates, as determined by Kaplan-Meierestimates.

In one embodiment, provided herein are methods for preventing ordelaying a Response Evaluation Criteria in Solid Tumors (RECIST 1.1) ofprogressive disease in a patient, comprising administering an effectiveamount of a solid form of Compound 1 described herein to a patienthaving a solid tumor as described herein. In one embodiment theprevention or delaying of progressive disease is characterized orachieved by a change in overall size of the target lesions, of forexample, between −30% and +20% compared to pre-treatment. In anotherembodiment, the change in size of the target lesions is a reduction inoverall size of more than 30%, for example, more than 50% reduction intarget lesion size compared to pre-treatment. In another, the preventionis characterized or achieved by a reduction in size or a delay inprogression of non-target lesions compared to pre-treatment. In oneembodiment, the prevention is achieved or characterized by a reductionin the number of target lesions compared to pre-treatment. In another,the prevention is achieved or characterized by a reduction in the numberor quality of non-target lesions compared to pre-treatment. In oneembodiment, the prevention is achieved or characterized by the absenceor the disappearance of target lesions compared to pre-treatment. Inanother, the prevention is achieved or characterized by the absence orthe disappearance of non-target lesions compared to pre-treatment. Inanother embodiment, the prevention is achieved or characterized by theprevention of new lesions compared to pre-treatment. In yet anotherembodiment, the prevention is achieved or characterized by theprevention of clinical signs or symptoms of disease progression comparedto pre-treatment, such as cancer-related cachexia or increased pain. Inone embodiment, the cases is a cancer set forth in Table 44.

In certain embodiments, provided herein are methods for decreasing thesize of target lesions in a patient compared to pre-treatment,comprising administering an effective amount of a solid form of Compound1 described herein to a patient having a cancer, in particular a solidtumor as described herein.

In certain embodiments, provided herein are methods for decreasing thesize of a non-target lesion in a patient compared to pre-treatment,comprising administering an effective amount of a solid form of Compound1 described herein to a patient having a cancer, in particular a solidtumor as described herein.

In certain embodiments, provided herein are methods for achieving areduction in the number of target lesions in a patient compared topre-treatment, comprising administering an effective amount of a solidform of Compound 1 described herein to a patient having a cancer, inparticular a solid tumor as described herein.

In certain embodiments, provided herein are methods for achieving areduction in the number of non-target lesions in a patient compared topre-treatment, comprising administering an effective amount a solid formof Compound 1 described herein to a patient having a cancer, inparticular a solid tumor as described herein.

In certain embodiments, provided herein are methods for achieving adisappearance of all target lesions in a patient, comprisingadministering an effective amount of a solid form of Compound 1described herein to a patient having a cancer, in particular a solidtumor as described herein.

In certain embodiments, provided herein are methods for achieving adisappearance of all non-target lesions in a patient, comprisingadministering an effective amount of a solid form of Compound 1described herein to a patient having a cancer, in particular a solidtumor as described herein.

In certain embodiments, provided herein are methods for treating acancer, in particular a solid tumor as described herein, the methodscomprising administering an effective amount of a solid form of Compound1 described herein to a patient having a cancer, in particular a solidtumor, wherein the treatment results in a complete response, partialresponse or stable disease, as determined by Response EvaluationCriteria in Solid Tumors (RECIST 1.1).

In certain embodiments, provided herein are methods for treating acancer, in particular a solid tumor as described herein, the methodscomprising administering an effective amount of a solid form of Compound1 described herein to a patient having a cancer, in particular a solidtumor as described herein, wherein the treatment results in a reductionin target lesion size, a reduction in non-target lesion size and/or theabsence of new target and/or non-target lesions, compared topre-treatment. In one embodiment, the cases is a cancer set forth inTable 44.

In certain embodiments, provided herein are methods for treating acancer, in particular a solid tumor as described herein, the methodscomprising administering an effective amount a solid form of Compound 1described herein to a patient having a cancer, in particular a solidtumor as described herein, wherein the treatment results in preventionor retarding of clinical progression, such as cancer-related cachexia orincreased pain.

In another embodiment, provided herein are methods for inducing atherapeutic response characterized with the International WorkshopCriteria (IWC) for NHL (see Cheson B D, Pfistner B, Juweid, M E, et. al.Revised Response Criteria for Malignant Lymphoma. J. Clin. Oncol: 2007:(25) 579-586) of a patient, comprising administering an effective amounta solid form of Compound 1 described herein to a patient having acancer, in particular hematological cancers such as lymphoma, asdescribed herein. In another embodiment, provided herein are methods forachieving complete remission, partial remission or stable disease, asdetermined by the International Workshop Criteria (IWC) for NHL in apatient, comprising administering an effective amount of a solid form ofCompound 1 described herein to a patient having a cancer, in particularhematological cancers such as lymphoma, as described herein. In anotherembodiment, provided herein are methods for achieving an increase inoverall survival, progression-free survival, event-free survival, timeto progression, disease-free survival or lymphoma-free survival asdetermined by the International Workshop Criteria (IWC) for NHL in apatient, comprising administering an effective amount of a solid form ofCompound 1 described herein to a patient having a cancer, in particularhematological cancers such as lymphoma, as described herein.

In another embodiment, provided herein are methods for inducing atherapeutic response assessed with the International Uniform ResponseCriteria for Multiple Myeloma (IURC) (see Durie B G M, Harousseau J-L,Miguel J S, et al. International uniform response criteria for multiplemyeloma. Leukemia, 2006; (10) 10: 1-7) of a patient, comprisingadministering an effective amount of a solid form of Compound 1 to apatient having a cancer, in particular multiple myeloma. In anotherembodiment, provided herein are methods for achieving a stringentcomplete response, complete response, very good partial response, orpartial response, as determined by the International Uniform ResponseCriteria for Multiple Myeloma (IURC) in a patient, comprisingadministering an effective amount of a solid form of Compound 1described herein to a patient having a cancer, in particular multiplemyeloma. In another embodiment, provided herein are methods forachieving an increase in overall survival, progression-free survival,event-free survival, time to progression, or disease-free survival in apatient, comprising administering an effective amount of a solid form ofCompound 1 described herein to a patient having a cancer, in particularmultiple myeloma.

In another embodiment, provided herein are methods for inducing atherapeutic response assessed with the Response Assessment forNeuro-Oncology (RANO) Working Group for GBM (see Wen P., Macdonald, DR., Reardon, D A., et al. Updated response assessment criteria forhigh-grade gliomas: Response assessment in neuro-oncology working group.J. Clin. Oncol. 2010; 28: 1963-1972) of a patient, comprisingadministering an effective amount of a solid form of Compound 1described herein to a patient having a cancer, in particularglioblastoma multiforme (GBM). In one embodiment, RANO will be used toestablish the proportion of subjects progression-free at 6 months fromDay 1 of treatment relative to efficacy evaluable subjects in the GBMtype.

In another embodiment, provided herein are methods for improving theEastern Cooperative Oncology Group Performance Status (ECOG) of apatient, comprising administering an effective amount a solid form ofCompound 1 described herein to a patient having a cancer, in particulara solid tumor or hematological cancer as described herein.

In another embodiment, provided herein are methods for inducing atherapeutic response assessed by Positron Emission Tomography (PET)outcome of a patient, comprising administering an effective amount of asolid form of Compound 1 described herein to a patient having a cancer,in particular a solid tumor or hematological cancer as described herein.In certain embodiments, provided herein are methods for treating acancer, in particular a solid tumor or hematological cancer as describedherein, the methods comprising administering an effective amount of asolid form of Compound 1 described herein to a patient having a cancer,in particular a solid tumor or hematological cancer as described herein,wherein the treatment results in a reduction in tumor metabolicactivity, for example, as measured by PET imaging.

Further provided herein are methods for treating patients who have beenpreviously treated for a cancer, in particular a solid tumor or ahematological cancer as described herein, as well as those who have notpreviously been treated. Such methods include administration of a solidform of Compound 1 described herein. Because patients with a cancer haveheterogeneous clinical manifestations and varying clinical outcomes, thetreatment given to a patient may vary, depending on his/her prognosis.The skilled clinician will be able to readily determine without undueexperimentation specific secondary agents, types of surgery, and typesof non-drug based standard therapy that can be effectively used to treatan individual patient with a cancer.

Biomarkers

In one embodiment, provided herein are methods for modulating the levelsof a biomarker in a subject having a cancer as described herein,comprising administering an effective amount of a solid form of Compound1 described herein, to said subject. In some such embodiments, themodulation of the biomarker is assessed in a biological sample of thesubject, such as in circulating blood, skin biopsies, tumor biopsies,circulating tumor cells, hair, and/or urine. In one embodiment, thebiological sample is peripheral blood mononuclear cells (PBMC). In suchembodiments, the amount of biomarker modulation is assessed bycomparison of the amount of biomarker before and after administration ofthe solid form of Compound 1 described herein or pharmaceuticalcomposition thereof. In some embodiments, the modulation in biomarker isa reduction of about 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%,90%, 95%, 99%, or about 100% compared to baseline levels. In some otherembodiments, the modulation in biomarker is an increase of about 10%,20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, 99%, or about100% compared to baseline levels.

In some embodiments, the biomarker is ERK, RSK1, DUSP4, DUSP5, DUSP6,BMF, EFNA1, EGR1, ETV5, FOS, FOSL1, GJA1, IL-8, cMyc, Cyclin D1, YAP,SPRY2, SPRY4, Axin2, CTGF, AREG, CYR61, CXCL1, HAS2, HES1, MAFF, CITED2,ELF3, or PD-Ll. In some such embodiments, the modulation is measured bymeasurement of the reduction of phosphorylation levels of one or more ofERK and RSK1. In some embodiments, the modulation in biomarker is areduction of about 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%,90%, 95%, 99%, or about 100% compared to baseline levels. In some otherembodiments, the modulation in biomarker is an increase of about 10%,20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, 99%, or about100% compared to baseline levels.

In some embodiments, the biomarker is one or more of DUSP4, DUSP6,cyclin D1, c-Myc, SPRY2, and YAP. In some such embodiments, themodulation is measured by measurement of the reduction in mRNA and/orprotein expression levels of one or more of DUSP4, DUSP6, cyclin D1,c-Myc, and YAP. In some such embodiments, the modulation is measured bymeasurement of the reduction in mRNA and/or protein expression levels ofone or more of DUSP4, DUSP6, SPRY2, c-Myc and cyclin D1. In someembodiments, the modulation in biomarker is a reduction of about 10%,20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, 99%, or about100% compared to baseline levels.

In some embodiments, the biomarker is one or more of DUSP4, DUSP6,cyclin D1, c-Myc, SPRY2, and YAP. In some such embodiments, themodulation is measured by measurement of the reduction in mRNA and/orprotein expression levels of one or more of DUSP4, DUSP6, cyclin D1,c-Myc, and YAP. In some such embodiments, the modulation is measured bymeasurement of the reduction in mRNA and/or protein expression levels ofone or more of DUSP4, DUSP6, SPRY2, c-Myc and cyclin D1. In someembodiments, the modulation in biomarker is a reduction of about 10%,20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, 99%, or about100% compared to baseline levels.

In some embodiments, the biomarker is one or more of DUSP5, DUSP6, EGR1,ETV5, FOS, FOSL1, IL8, SPRY2, and SPRY4. In some such embodiments, themodulation is measured by measurement of the reduction in mRNA and/orprotein expression levels of one or more of DUSP5, DUSP6, EGR1, ETV5,FOS, FOSL1, IL8, SPRY2, and SPRY4. In some embodiments, the modulationin biomarker is a reduction of about 10%, 20%, 25%, 30%, 40%, 50%, 60%,70%, 75%, 80%, 90%, 95%, 99%, or about 100% compared to baseline levels.

In some embodiments, the biomarker is one or more of BMF and EFNA. Insome such embodiments, the modulation is measured by measurement of theincrease in mRNA and/or protein expression levels of one or more of BMFand EFNA1. In some embodiments, the modulation in biomarker is anincrease of about 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%,95%, 99%, or about 100% compared to baseline levels.

In some embodiments, the biomarker is GJA1. In some such embodiments,the modulation is measured by measurement of the modulation in mRNAand/or protein expression levels of one or more of GJA1. In some suchembodiments, the modulation in biomarker is a reduction of about 10%,20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, 99%, or about100% compared to baseline levels. In some embodiments, the modulation inbiomarker is an increase of about 10%, 20%, 25%, 30%, 40%, 50%, 60%,70%, 75%, 80%, 90%, 95%, 99%, or about 100% compared to baseline levels.

In some embodiments, the biomarker is one or more of Axin2, CTGF, Cur61and AREG. In some such embodiments, the modulation is measured bymeasurement of the reduction in mRNA and/or protein expression levels ofone or more of Axin2, CTGF, and AREG. In some embodiments, themodulation in biomarker is a reduction of about 10%, 20%, 25%, 30%, 40%,50%, 60%, 70%, 75%, 80%, 90%, 95%, 99%, or about 100% compared tobaseline levels.

In some embodiments, the biomarker is one or more of CYR61, CXCL1, HAS2,HES1 and MAFF. In some such embodiments, the modulation is measured bymeasurement of the reduction in mRNA and/or protein expression levels ofone or more of CYR61, CXCL1, HAS2, HES1 and MAFF. In some embodiments,the modulation in biomarker is a reduction of about 10%, 20%, 25%, 30%,40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, 99%, or about 100% compared tobaseline levels.

In some embodiments, the biomarker is one or more of CITED2 and ELF3. Insome such embodiments, the modulation is measured by measurement of theincrease in mRNA and/or protein expression levels of one or more ofCITED2 and ELF3. In some embodiments, the modulation in biomarker is anincrease of about 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%,95%, 99%, or about 100% compared to baseline levels.

In some embodiments, the biomarker is PD-L1. In some embodiments, themodulation in the levels of biomarker is a reduction in cell surfaceexpression levels of PD-L1. In some embodiments, the modulation inbiomarker is a reduction of about 10%, 20%, 25%, 30%, 40%, 50%, 60%,70%, 75%, 80%, 90%, 95%, 99%, or about 100% compared to baseline levels.

In another embodiment, the biomarker is IFNγ or IL-2. In some suchembodiments, the modulation in the levels of biomarker is an increase inmRNA and/or protein expression levels of IFNγ or IL-2. In some suchembodiments, the modulation in mRNA and/or protein expression levels ofIFNγ or IL-2 is an increase of about 10%, 20%, 25%, 30%, 40%, 50%, 60%,70%, 75%, 80%, 90%, 95%, 99%, or about 100% compared to baseline levels.

In another embodiment, the biomarker is IL-8. In some such embodiments,the modulation in the levels of biomarker is a decrease in mRNA and/orprotein expression levels of IL-8. In some such embodiments, themodulation in mRNA and/or protein expression levels of IL-8 is andecrease of about 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%,95%, 99%, or about 100% compared to baseline levels.

In one embodiment, provided herein are methods for inhibitingphosphorylation of ERK and/or RSK1 in a subject having a cancer asdescribed herein, comprising administering an effective amount of asolid form of Compound 1 as described herein to said subject. In somesuch embodiments, the inhibition of phosphorylation is assessed in abiological sample of the subject, such as in circulating blood and/ortumor cells, skin biopsies and/or tumor biopsies or aspirate. In suchembodiments, the amount of inhibition of phosphorylation is assessed bycomparison of the amount of phospho-ERK and/or RSK1 before and afteradministration of the solid form of Compound 1 provided herein. Incertain embodiments, provided herein are methods for measuringinhibition of phosphorylation of ERK and/or RSK1, in a subject having acancer as described herein, comprising administering an effective amountof a solid form of Compound 1 provided herein to said subject, measuringthe amount of phosphorylated ERK and/or RSK1 in said subject, andcomparing said amount of phosphorylated ERK and/or RSK to that of saidsubject prior to administration of an effective amount of the solid formof Compound 1 provided herein. In some embodiments, the biologicalsample is a tumor biopsy. In another embodiment, the biological sampleis PBMC. In still another embodiment, the biological sample iscirculating tumor cells.

In certain embodiments, provided herein are methods for inhibitingphosphorylation of ERK and/or RSK1 in a biological sample of a subjecthaving a cancer as described herein, comprising administering aneffective amount of a solid form of Compound 1 provided herein to saidsubject and comparing the amount of phosphorylated ERK and/or RSK1 in abiological sample of a subject obtained prior to and afteradministration of said solid form of Compound 1 provided herein, whereinless phosphorylated ERK and/or RSK1 in said biological sample obtainedafter administration of said solid form of Compound 1 provided hereinrelative to the amount of phosphorylated ERK and/or RSK1 in saidbiological sample obtained prior to administration of said solid form ofCompound 1 provided herein indicates inhibition. In some embodiments,the biological sample is a tumor biopsy. In another embodiment, thebiological sample is PBMC. In still another embodiment, the biologicalsample is circulating tumor cells.

Further provided herein are methods for determining whether a patient issensitive to a solid form of Compound 1 described herein, comprisingadministering said patient said solid form of Compound 1 describedherein and determining whether or not ERK and/or RSK1 phosphorylation isinhibited in said patient by measuring the amount of phosphorylated ERKand/or RSK1 in a biological sample from said patient prior to and afterthe administration of a solid form of Compound 1 described herein tosaid patient, wherein inhibition of ERK and/or RSK1 phosphorylationindicates that said patient is sensitive to said solid form of Compound1 described herein. In some such embodiments, the method additionallycomprises administering an effective amount of a solid form of Compound1, as described herein. In some embodiments, the biological sample is atumor biopsy. In another embodiment, the biological sample is PBMC. Instill another embodiment, the biological sample is circulating tumorcells.

Further provided herein are methods for determining the effective amountof a solid form of Compound 1 described herein for the treatment of acancer treatable by inhibition of phosphorylation of ERK and/or RSK1 ina patient, comprising administering said patient varying doses of said asolid form of Compound 1 described herein and determining the amount ofERK and/or RSK1 phosphorylation inhibition in said patient resultingfrom each dose of said a solid form of Compound 1 described herein bymeasuring the amount of phosphorylated ERK and/or RSK1 in a biologicalsample from said patient prior to and after the administration of eachdose of a solid form of Compound 1 described herein to said patient,wherein inhibition of ERK and/or RSK1 phosphorylation by at least about10%, about 20%, about 30%, about 40%, about 50% or greater than about50%, corresponds to an effective amount of a solid form of Compound 1described herein. In some such embodiments, the method additionallycomprises administering an effective amount of a solid form of Compound1, as described herein. In some embodiments, the biological sample is atumor biopsy. In another embodiment, the biological sample is PBMC. Instill another embodiment, the biological sample is circulating tumorcells.

Further provided herein are methods for predicting response to treatmentwith a solid form of Compound 1 described herein in a patient having acancer, the method comprising: a) obtaining a biological test samplefrom the patient's cancer; b) obtaining the mRNA and/or proteinexpression levels of one or more of DUSP4, DUSP5, DUSP6, EGR1, ETV5,FOS, FOSL1, IL-8, cMyc, Cyclin D1, YAP, SPRY2, SPRY4, Axin2, CTGF, AREG,CYR61, CXCL1, HAS2, HES1, and MAFF in said biological test sample; c)comparing said mRNA and/or protein expression levels to the mRNA and/orprotein expression levels of a biological wild-type sample; wherein areduction in mRNA and/or protein expression levels in said patient'sbiological test sample relative to said biological wild-type sample,indicates an increased likelihood of response to treatment with a solidform of Compound 1 described herein of said patient's cancer. In somesuch embodiments, the method additionally comprises administering aneffective amount of a solid form of Compound 1, as described herein. Insome embodiments, the biological sample is a tumor biopsy. In anotherembodiment, the biological sample is PBMC. In still another embodiment,the biological sample is circulating tumor cells.

Further provided herein are methods for predicting therapeutic efficacyof treatment with a solid form of Compound 1 described herein of apatient having a cancer, the method comprising: a) obtaining abiological test sample from the patient's cancer; b) obtaining the mRNAand/or protein expression levels of one or more of DUSP4, DUSP5, DUSP6,EGR1, ETV5, FOS, FOSL1, IL-8, cMyc, Cyclin D1, YAP, SPRY2, SPRY4, Axin2,CTGF, AREG, CYR61, CXCL1, HAS2, HES1, and MAFF in said biological testsample; c) comparing said mRNA and/or protein expression levels to themRNA and/or protein expression levels of a biological wild-type sample;wherein a reduction in mRNA and/or protein expression levels indicatesan increased likelihood of therapeutic efficacy of said treatment with asolid form of Compound 1 described herein for said patient. In some suchembodiments, the method additionally comprises administering aneffective amount of a solid form of Compound 1, as described herein. Insome embodiments, the biological sample is a tumor biopsy. In anotherembodiment, the biological sample is PBMC. In still another embodiment,the biological sample is circulating tumor cells.

Further provided herein are methods for determining whether a patient issensitive to a solid form of Compound 1 described herein, comprisingadministering said patient said solid form of Compound 1 describedherein and determining whether or not mRNA and/or protein expressionlevels of one or more of DUSP4, DUSP5, DUSP6, EGR1, ETV5, FOS, FOSL1,IL-8, cMyc, Cyclin D1, YAP, SPRY2, SPRY4, Axin2, CTGF, AREG, CYR61,CXCL1, HAS2, HES1, and MAFF, are inhibited in said patient, by measuringthe amount of mRNA and/or protein expression levels of one or more ofDUSP4, DUSP5, DUSP6, EGR1, ETV5, FOS, FOSL1, IL-8, cMyc, Cyclin D1, YAP,SPRY2, SPRY4, Axin2, CTGF, AREG, CYR61, CXCL1, HAS2, HES1, and MAFF in abiological sample from said patient, prior to and after theadministration of a solid form of Compound 1 described herein to saidpatient. In some such embodiments, the method additionally comprisesadministering an effective amount of a solid form of Compound 1, asdescribed herein. In some embodiments, the biological sample is a tumorbiopsy. In another embodiment, the biological sample is PBMC. In stillanother embodiment, the biological sample is circulating tumor cells.

Further provided herein are methods for determining the effective amountof a solid form of Compound 1 described herein for the treatment of acancer treatable by inhibition of mRNA and/or protein expression levelsof one or more of DUSP4, DUSP5, DUSP6, EGR1, ETV5, FOS, FOSL1, IL-8,cMyc, Cyclin D1, YAP, SPRY2, SPRY4, Axin2, CTGF, AREG, CYR61, CXCL1,HAS2, HES1, and MAFF in a patient, comprising administering said patientvarying doses of said solid form of Compound 1 described herein anddetermining the amount of mRNA and/or protein expression levels of oneor more of DUSP4, DUSP5, DUSP6, EGR1, ETV5, FOS, FOSL1, IL-8, cMyc,Cyclin D1, YAP, SPRY2, SPRY4, Axin2, CTGF, AREG, CYR61, CXCL1, HAS2,HES1, and MAFF inhibition in said patient, resulting from each dose ofsaid solid form of Compound 1 described herein by measuring the amountof mRNA and/or protein expression levels of one or more of DUSP4, DUSP5,DUSP6, EGR1, ETV5, FOS, FOSL1, IL-8, cMyc, Cyclin D1, YAP, SPRY2, SPRY4,Axin2, CTGF, AREG, CYR61, CXCL1, HAS2, HES1, and MAFF in a biologicalsample from said patient, prior to and after the administration of eachdose of a solid form of Compound 1 described herein to said patient. Insome such embodiments, the method additionally comprises administeringan effective amount of a solid form of Compound 1, as described herein.In some embodiments, the biological sample is a tumor biopsy. In anotherembodiment, the biological sample is PBMC. In still another embodiment,the biological sample is circulating tumor cells.

Further provided herein are methods for predicting response to treatmentwith a solid form of Compound 1 described herein in a patient having acancer, the method comprising: a) obtaining a biological test samplefrom the patient's cancer; b) obtaining the mRNA and/or proteinexpression levels of one or more of BMF, EFNA1, CITED2, and ELF3 in saidbiological test sample; c) comparing said mRNA and/or protein expressionlevels to the mRNA and/or protein expression levels of a biologicalwild-type sample; wherein an increase in mRNA and/or protein expressionlevels in said patient's biological test sample relative to saidbiological wild-type sample, indicates an increased likelihood ofresponse to treatment with a solid form of Compound 1 described hereinof said patient's cancer. In some such embodiments, the methodadditionally comprises administering an effective amount of a solid formof Compound 1, as described herein. In some embodiments, the biologicalsample is a tumor biopsy. In another embodiment, the biological sampleis PBMC. In still another embodiment, the biological sample iscirculating tumor cells.

Further provided herein are methods for predicting therapeutic efficacyof treatment with a solid form of Compound 1 described herein of apatient having a cancer, the method comprising: a) obtaining abiological test sample from the patient's cancer; b) obtaining the mRNAand/or protein expression levels of one or more of BMF, EFNA1, CITED2,and ELF3 in said biological test sample; c) comparing said mRNA and/orprotein expression levels to the mRNA and/or protein expression levelsof a biological wild-type sample; wherein an increase in mRNA and/orprotein expression levels indicates an increased likelihood oftherapeutic efficacy of said solid form of Compound 1 described hereintreatment for said patient. In some such embodiments, the methodadditionally comprises administering an effective amount of a solid formof Compound 1, as described herein. In some embodiments, the biologicalsample is a tumor biopsy. In another embodiment, the biological sampleis PBMC. In still another embodiment, the biological sample iscirculating tumor cells.

Further provided herein are methods for determining whether a patient issensitive to a solid form of Compound 1 described herein, comprisingadministering said patient said solid form of Compound 1 describedherein and determining whether or not mRNA and/or protein expressionlevels of one or more of BMF, EFNA1, CITED2, and ELF3 are increased insaid patient, by measuring the amount of mRNA and/or protein expressionlevels of one or more of BMF, EFNA1, CITED2, and ELF3 in a biologicalsample from said patient, prior to and after the administration of asolid form of Compound 1 described herein to said patient. In some suchembodiments, the method additionally comprises administering aneffective amount of a solid form of Compound 1, as described herein. Insome embodiments, the biological sample is a tumor biopsy. In anotherembodiment, the biological sample is PBMC. In still another embodiment,the biological sample is circulating tumor cells.

Further provided herein are methods for determining the effective amountof a solid form of Compound 1 described herein for the treatment of acancer treatable by an increase of mRNA and/or protein expression levelsof one or more of BMF, EFNA1, CITED2, and ELF3 in a patient, comprisingadministering said patient varying doses of said solid form of Compound1 described herein, and determining the amount of mRNA and/or proteinexpression levels of one or more of BMF, EFNA1, CITED2, and ELF3increase in said patient resulting from each dose of said solid form ofCompound 1 described herein by measuring the amount of mRNA and/orprotein expression levels of one or more of BMF, EFNA1, CITED2, and ELF3in a biological sample from said patient, prior to and after theadministration of each dose of a solid form of Compound 1 describedherein to said patient. In some such embodiments, the methodadditionally comprises administering an effective amount of a solid formof Compound 1, as described herein. In some embodiments, the biologicalsample is a tumor biopsy. In another embodiment, the biological sampleis PBMC. In still another embodiment, the biological sample iscirculating tumor cells.

Further provided herein are methods for predicting response to treatmentwith a solid form of Compound 1 described herein in a patient having acancer, the method comprising: a) obtaining a biological test samplefrom the patient's cancer; b) obtaining the mRNA and/or proteinexpression levels of GJA1 in said biological test sample; c) comparingsaid mRNA and/or protein expression levels to the mRNA and/or proteinexpression levels of a biological wild-type sample; wherein a reductionin mRNA and/or protein expression levels in said patient's biologicaltest sample relative to said biological wild-type sample, indicates anincreased likelihood of response to treatment with a solid form ofCompound 1 described herein of said patient's cancer. In some suchembodiments, the method additionally comprises administering aneffective amount of a solid form of Compound 1, as described herein. Insome embodiments, the biological sample is a tumor biopsy. In anotherembodiment, the biological sample is PBMC. In still another embodiment,the biological sample is circulating tumor cells.

Further provided herein are methods for predicting therapeutic efficacyof treatment with a solid form of Compound 1 described herein of apatient having a cancer, the method comprising: a) obtaining abiological test sample from the patient's cancer; b) obtaining the mRNAand/or protein expression levels of GJA1 in said biological test sample;c) comparing said mRNA and/or protein expression levels to the mRNAand/or protein expression levels of a biological wild-type sample;wherein a reduction in mRNA and/or protein expression levels indicatesan increased likelihood of therapeutic efficacy of said treatment with asolid form of Compound 1 described herein for said patient. In some suchembodiments, the method additionally comprises administering aneffective amount of a solid form of Compound 1, as described herein. Insome embodiments, the biological sample is a tumor biopsy. In anotherembodiment, the biological sample is PBMC. In still another embodiment,the biological sample is circulating tumor cells.

Further provided herein are methods for determining whether a patient issensitive to a solid form of Compound 1 described herein, comprisingadministering said patient said solid form of Compound 1 describedherein and determining whether or not mRNA and/or protein expressionlevels of GJA1 are inhibited in said patient, by measuring the amount ofmRNA and/or protein expression levels of GJA1 in a biological samplefrom said patient, prior to and after the administration of a solid formof Compound 1 described herein to said patient. In some suchembodiments, the method additionally comprises administering aneffective amount of a solid form of Compound 1, as described herein. Insome embodiments, the biological sample is a tumor biopsy. In anotherembodiment, the biological sample is PBMC. In still another embodiment,the biological sample is circulating tumor cells.

Further provided herein are methods for determining the effective amountof a solid form of Compound 1 described herein for the treatment of acancer treatable by inhibition of mRNA and/or protein expression levelsof GJA1 in a patient, comprising administering said patient varyingdoses of said solid form of Compound 1 described herein and determiningthe amount of mRNA and/or protein expression levels of GJA1 inhibitionin said patient, resulting from each dose of said solid form of Compound1 described herein by measuring the amount of mRNA and/or proteinexpression levels of GJA1 in a biological sample from said patient,prior to and after the administration of each dose of a solid form ofCompound 1 described herein to said patient. In some such embodiments,the method additionally comprises administering an effective amount of asolid form of Compound 1, as described herein. In some embodiments, thebiological sample is a tumor biopsy. In another embodiment, thebiological sample is PBMC. In still another embodiment, the biologicalsample is circulating tumor cells.

Further provided herein are methods for predicting response to treatmentwith a solid form of Compound 1 described herein in a patient having acancer, the method comprising: a) obtaining a biological test samplefrom the patient's cancer; b) obtaining the mRNA and/or proteinexpression levels of GJA1 in said biological test sample; c) comparingsaid mRNA and/or protein expression levels to the mRNA and/or proteinexpression levels of a biological wild-type sample; wherein an increasein mRNA and/or protein expression levels in said patient's biologicaltest sample relative to said biological wild-type sample, indicates anincreased likelihood of response to a solid form of Compound 1 describedherein treatment of said patient's cancer. In some such embodiments, themethod additionally comprises administering an effective amount of asolid form of Compound 1, as described herein. In some embodiments, thebiological sample is a tumor biopsy. In another embodiment, thebiological sample is PBMC. In still another embodiment, the biologicalsample is circulating tumor cells.

Further provided herein are methods for predicting therapeutic efficacyof treatment with a solid form of Compound 1 described herein of apatient having a cancer, the method comprising: a) obtaining abiological test sample from the patient's cancer; b) obtaining the mRNAand/or protein expression levels of GJA1 in said biological test sample;c) comparing said mRNA and/or protein expression levels to the mRNAand/or protein expression levels of a biological wild-type sample;wherein an increase in mRNA and/or protein expression levels indicatesan increased likelihood of therapeutic efficacy of said treatment with asolid form of Compound 1 described herein for said patient. In some suchembodiments, the method additionally comprises administering aneffective amount of a solid form of Compound 1, as described herein. Insome embodiments, the biological sample is a tumor biopsy. In anotherembodiment, the biological sample is PBMC. In still another embodiment,the biological sample is circulating tumor cells.

Further provided herein are methods for determining whether a patient issensitive to a solid form of Compound 1, comprising administering saidpatient said a solid form of Compound 1 described herein and determiningwhether or not mRNA and/or protein expression levels of GJA1 areincreased in said patient, by measuring the amount of mRNA and/orprotein expression levels of GJA1 in a biological sample from saidpatient, prior to and after the administration of a solid form ofCompound 1 described herein to said patient. In some such embodiments,the method additionally comprises administering an effective amount of asolid form of Compound 1, as described herein. In some embodiments, thebiological sample is a tumor biopsy. In another embodiment, thebiological sample is PBMC. In still another embodiment, the biologicalsample is circulating tumor cells.

Further provided herein are methods for determining the effective amountof a solid form of Compound 1 for the treatment of a cancer treatable byan increase of mRNA and/or protein expression levels of GJA1 in apatient, comprising administering said patient varying doses of saidsolid form of Compound 1 described herein, and determining the amount ofmRNA and/or protein expression levels of GJA1 increase in said patientresulting from each dose of said solid form of Compound 1 describedherein by measuring the amount of mRNA and/or protein expression levelsof GJA1 in a biological sample from said patient, prior to and after theadministration of each dose of a solid form of Compound 1 describedherein to said patient. In some such embodiments, the methodadditionally comprises administering an effective amount of a solid formof Compound 1, as described herein. In some embodiments, the biologicalsample is a tumor biopsy. In another embodiment, the biological sampleis PBMC. In still another embodiment, the biological sample iscirculating tumor cells.

Further provided herein are methods for predicting response to treatmentwith a solid form of Compound 1 described herein in a patient having acancer, the method comprising: a) obtaining a biological test samplefrom the patient's cancer; b) obtaining the cell surface expressionlevels of PD-L1 in said biological test sample; c) comparing said cellsurface expression levels of PD-L1 to the cell surface expression levelsof PD-L1 of a biological wild-type sample; wherein a reduction in cellsurface expression levels of PD-L1 indicates an increased likelihood ofresponse to a solid form of Compound 1 described herein treatment ofsaid patient's cancer. In some such embodiments, the method additionallycomprises administering an effective amount of a solid form of Compound1, as described herein. In some embodiments, the biological sample is atumor biopsy. In another embodiment, the biological sample is PBMC. Instill another embodiment, the biological sample is circulating tumorcells.

Further provided herein are methods for predicting therapeutic efficacyof treatment with a solid form of Compound 1 described herein of apatient having a cancer, the method comprising: a) obtaining abiological test sample from the patient's cancer; b) obtaining the cellsurface expression levels of PD-L1 in said biological test sample; c)comparing said cell surface expression levels of PD-L1 to the cellsurface expression levels of PD-L1 of a biological wild-type sample;wherein a reduction in cell surface expression levels of PD-L1 indicatesan increased likelihood of therapeutic efficacy of said treatment with asolid form of Compound 1 described herein for said patient. In some suchembodiments, the method additionally comprises administering aneffective amount of a solid form of Compound 1, as described herein. Insome embodiments, the biological sample is a tumor biopsy. In anotherembodiment, the biological sample is PBMC. In still another embodiment,the biological sample is circulating tumor cells.

Further provided herein are methods for determining whether a patient issensitive to a solid form of Compound 1, comprising administering saidpatient said a solid form of Compound 1 described herein and determiningwhether or not cell surface expression levels of PD-L1 are inhibited insaid patient by measuring the amount of cell surface expression levelsof PD-L1 in a biological sample from said patient prior to and after theadministration of a solid form of Compound 1 described herein to saidpatient. In some such embodiments, the method additionally comprisesadministering an effective amount of a solid form of Compound 1, asdescribed herein. In some embodiments, the biological sample is a tumorbiopsy. In another embodiment, the biological sample is PBMC. In stillanother embodiment, the biological sample is circulating tumor cells.

Further provided herein are methods for determining the effective amountof a solid form of Compound 1 described herein for the treatment of acancer treatable by cell surface expression levels of PD-L1 in apatient, comprising administering said patient varying doses of saidsolid form of Compound 1 described herein and determining the amount ofcell surface expression levels of PD-L1 inhibition in said patientresulting from each dose of said solid form of Compound 1 describedherein by measuring the amount of cell surface expression levels ofPD-L1 in a biological sample from said patient prior to and after theadministration of each dose of a solid form of Compound 1 describedherein to said patient. In some such embodiments, the methodadditionally comprises administering an effective amount of a solid formof Compound 1, as described herein. In some embodiments, the biologicalsample is a tumor biopsy. In another embodiment, the biological sampleis PBMC. In still another embodiment, the biological sample iscirculating tumor cells.

Combination Therapy

Solid forms of Compound 1 provided herein can also be combined or usedin combination with other therapeutic agents useful in the treatmentand/or prevention of cancer described herein.

In one embodiment, provided herein is a method of treating, preventing,or managing cancer, comprising administering to a patient a solid formof Compound 1 provided herein in combination with one or more secondactive agents, and optionally in combination with radiation therapy,blood transfusions, or surgery. Examples of second active agents aredisclosed herein.

As used herein, the term “in combination” includes the use of more thanone therapy (e.g., one or more prophylactic and/or therapeutic agents).However, the use of the term “in combination” does not restrict theorder in which therapies (e.g., prophylactic and/or therapeutic agents)are administered to a patient with a disease or disorder. A firsttherapy (e.g., a prophylactic or therapeutic agent such as a solid formof Compound 1 provided herein, can be administered prior to (e.g., 5minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before),concomitantly with, or subsequent to (e.g., 5 minutes, 15 minutes, 30minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks,5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of asecond therapy (e.g., a prophylactic or therapeutic agent) to thesubject. Triple therapy is also contemplated herein.

Administration of a solid form of Compound 1 provided herein and one ormore second active agents to a patient can occur simultaneously orsequentially by the same or different routes of administration. Thesuitability of a particular route of administration employed for aparticular active agent will depend on the active agent itself (e.g.,whether it can be administered orally without decomposing prior toentering the blood stream) and the cancer being treated.

The route of administration of a solid form of Compound 1 describedherein is independent of the route of administration of a secondtherapy. Thus, in accordance with these embodiments, a solid form ofCompound 1 described herein is administered intravenously, and thesecond therapy can be administered orally, parenterally,intraperitoneally, intravenously, intraarterially, transdermally,sublingually, intramuscularly, rectally, transbuccally, intranasally,liposomally, via inhalation, vaginally, intraoccularly, via localdelivery by catheter or stent, subcutaneously, intraadiposally,intraarticularly, intrathecally, or in a slow release dosage form. Inone embodiment, a solid form of Compound 1 described herein and a secondtherapy are administered by the same mode of administration, forexample, orally. In another embodiment, a solid form of Compound 1described herein is administered by one mode of administration, e.g.,orally, whereas the second agent (an anticancer agent) is administeredby another mode of administration, e.g., IV.

In one embodiment, the second active agent is administered, for example,orally, intravenously or subcutaneously, and once or twice daily in anamount of from about 1 to about 1000 mg, from about 5 to about 500 mg,from about 10 to about 350 mg, from about 50 to about 200 mg, from about1 to about 100 mg, from about 1 to about 200 mg, from about 1 to about300 mg, from about 1 to about 400 mg, or from about 1 to about 500 mg.The specific amount of the second active agent will depend on thespecific agent used, the type of disease being treated or managed, theseverity and stage of disease, and the amount of a solid form ofCompound 1 described herein described herein and any optional additionalactive agents concurrently administered to the patient.

One or more second active ingredients or agents can be used togetherwith a solid form of Compound 1 described herein in the methods andcompositions provided herein. Second active agents can be largemolecules (e.g., proteins) or small molecules (e.g., syntheticinorganic, organometallic, or organic molecules).

Examples of large molecule active agents include, but are not limitedto, hematopoietic growth factors, cytokines, and monoclonal andpolyclonal antibodies, particularly, therapeutic antibodies to cancerantigens. Typical large molecule active agents are biological molecules,such as naturally occurring or synthetic or recombinant proteins.Proteins that are particularly useful in the methods and compositionsprovided herein include proteins that stimulate the survival and/orproliferation of hematopoietic precursor cells lymphopoietic cells invitro or in vivo. Other useful proteins stimulate the division anddifferentiation of committed hematopoietic progenitors in cells in vitroor in vivo. Particular proteins include, but are not limited to:interleukins, such as IL-2 (including recombinant IL-2 (“rIL2”) andcanarypox IL-2), IL-10, IL-12, and IL-18; interferons, such asinterferon alfa-2a, interferon alfa-2b, interferon alfa-nl, interferonalfa-n3, interferon beta-Ia, and interferon gamma-I b; GM-CF and GM-CSF;and EPO.

In certain embodiments, GM-CSF, G-CSF, SCF or EPO is administeredsubcutaneously during about five days in a four or six week cycle in anamount ranging from about 1 to about 750 mg/m²/day, from about 25 toabout 500 mg/m²/day, from about 50 to about 250 mg/m²/day, or from about50 to about 200 mg/m²/day. In certain embodiments, GM-CSF may beadministered in an amount of from about 60 to about 500 mcg/m²intravenously over 2 hours or from about 5 to about 12 mcg/m²/daysubcutaneously. In certain embodiments, G-CSF may be administeredsubcutaneously in an amount of about 1 mcg/kg/day initially and can beadjusted depending on rise of total granulocyte counts. The maintenancedose of G-CSF may be administered in an amount of about 300 (in smallerpatients) or 480 mcg subcutaneously. In certain embodiments, EPO may beadministered subcutaneously in an amount of 10,000 Unit 3 times perweek.

Particular proteins that can be used in the methods and compositionsinclude, but are not limited to: filgrastim, sargramostim, andrecombinant EPO.

Recombinant and mutated forms of GM-CSF can be prepared as described inU.S. Pat. Nos. 5,391,485; 5,393,870; and 5,229,496; all of which areincorporated herein by reference. Recombinant and mutated forms of G-CSFcan be prepared as described in U.S. Pat. Nos. 4,810,643; 4,999,291;5,528,823; and 5,580,755; the entireties of which are incorporatedherein by reference.

Also provided for use in combination with a solid form of Compound 1described herein are native, naturally occurring, and recombinantproteins. Further encompassed are mutants and derivatives (e.g.,modified forms) of naturally occurring proteins that exhibit, in vivo,at least some of the pharmacological activity of the proteins upon whichthey are based. Examples of mutants include, but are not limited to,proteins that have one or more amino acid residues that differ from thecorresponding residues in the naturally occurring forms of the proteins.Also encompassed by the term “mutants” are proteins that lackcarbohydrate moieties normally present in their naturally occurringforms (e.g., nonglycosylated forms). Examples of derivatives include,but are not limited to, pegylated derivatives and fusion proteins, suchas proteins formed by fusing IgG1 or IgG3 to the protein or activeportion of the protein of interest. See, e.g., Penichet, M. L. andMorrison, S. L., J. Immunol. Methods 248:91-101 (2001).

Antibodies that can be used in combination with a solid form of Compound1 described herein include monoclonal and polyclonal antibodies.Examples of antibodies include, but are not limited to, trastuzumab,rituximab, bevacizumab, pertuzumab, tositumomab, edrecolomab, and G250.Solid forms of Compound 1 described herein can also be combined with, orused in combination with, anti-TNF-α antibodies, and/or anti-EGFRantibodies, such as, for example, cetuximab or panitumumab.

Antibodies that can be used in combination with a solid form of Compound1 described herein include immune checkpoint inhibitors, such as,anti-CTLA4, anti-PD1, anti-PD-L1, anti-Tim-3, anti-Lag-3 antibodies. Insome such embodiments, the PD-1 or PD-L1 antibodies are, for example,avelumab, durvalumab, MEDI0680, atezolizumab, BMS-936559, nivolumab,pembrolizumab, pidilizumab, or PDR-001. In one such embodiment, theanti-Lag-3 antibody is BMS-986016.

Additional antibodies that can be used in combination with a solid formof Compound 1 described herein include anti-RSPO antibodies.

Large molecule active agents may be administered in the form ofanti-cancer vaccines. For example, vaccines that secrete, or cause thesecretion of, cytokines such as IL-2, G-CSF, and GM-CSF can be used inthe methods and pharmaceutical compositions provided. See, e.g., Emens,L. A., et al., Curr. Opinion Mol. Ther. 3(1):77-84 (2001).

Second active agents that are small molecules can also be used toalleviate adverse effects associated with the administration of a solidform of Compound 1 described herein. However, like some large molecules,many are believed to be capable of providing an additive or synergisticeffect when administered with (e.g., before, after or simultaneously) asolid form of Compound 1 described herein. Examples of small moleculesecond active agents include, but are not limited to, anti-canceragents, antibiotics, immunosuppressive agents, and steroids.

In certain embodiments, the second agent is a BRAF inhibitor, an HSPinhibitor, a proteasome inhibitor, a FLT3 inhibitor, a MEK inhibitor, aPI3K inhibitor, an EGFR inhibitor, an immunomodulatory compound, or aTOR kinase inhibitor. In some such embodiments, the BRAF inhibitor issorafenib, dabrafenib, encorafenib, or vemurafenib. In some suchembodiment, the HSP inhibitor is geldanamycin, gamitrinib, luminespib,or radicicol. In some embodiments, the proteasome inhibitor isbortezomib, carfilzomib, ixazomib, disulfiram, oprozomib, delanzomib, orixazomib. In other embodiments, the FLT3 inhibitor is quizartinib,midostaurin, sorafenib, sunitinib, or lestaurtinib. In some suchembodiments, the MEK inhibitor is trametinib, cobimetinib, binimetinib,selumetinib, PD-325901, CI-1040 (PD184352) or TAK-733. In some otherembodiments, the PI3K inhibitor is AT7867, AZD 8055, BX-912,silmitasertib, pictilisib, MK-2206, or pilaralisib. In anotherembodiment, the EGFR inhibitor is gefitinib, erlotinib, afatinib,osimertinib (TAGRISSO), rociletinib, or lapatinib. In some otherembodiments, the TOR kinase inhibitor is CC-115, CC-223, OSI-027,AZD8055, sapanisertib, dactolisib, BGT226, voxtalisib (SAR-245409),apitolisib, omipalisib (GSK-2126458), PF-04691502, gedatolisib or PP242.In some embodiments, the immunomodulatory compound is thalidomide,lenalidomide, pomalidomide, CC-220, or CC-122.

Examples of additional anti-cancer agents to be used within the methodsor compositions described herein include, but are not limited to:acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin;aldesleukin; altretamine; ambomycin; ametantrone acetate; amsacrine;anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa;azotomycin; batimastat; benzodepa; bicalutamide; bisantrenehydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate;brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone;caracemide; carbetimer; carboplatin; carmustine; carubicinhydrochloride; carzelesin; cedefingol; celecoxib (COX-2 inhibitor);chlorambucil; cirolemycin; cisplatin; cladribine; clofarabine; crisnatolmesylate; cyclophosphamide; arabinoxylcytosine; dacarbazine; dabrafenib;dactinomycin; daunorubicin hydrochloride; decitabine; dexormaplatin;dezaguanine; dezaguanine mesylate; diaziquone; docetaxel; doxorubicin;doxorubicin hydrochloride; droloxifene; droloxifene citrate;dromostanolone propionate; duazomycin; edatrexate; eflornithinehydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine;epirubicin hydrochloride; erbulozole; esorubicin hydrochloride;estramustine; estramustine phosphate sodium; etanidazole; etoposide;etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine;fenretinide; floxuridine; fludarabine phosphate; fluorouracil;flurocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabinehydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide;ilmofosine; iproplatin; irinotecan; irinotecan hydrochloride; lanreotideacetate; letrozole; leuprolide acetate; liarozole hydrochloride;lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol;maytansine; mechlorethamine hydrochloride; megestrol acetate;melengestrol acetate; melphalan; menogaril; mercaptopurine;methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide;mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper;mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole;nogalamycin; omacetaxine; ormaplatin; oxisuran; paclitaxel; paclitaxelprotein-bound particles for injectable suspension, albumin bound(ABRAXANE®); pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; safingol; safingol hydrochloride; semustine;simtrazene; sorafenib; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; docetaxel; tegafur;teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;vemurafenib; verteporfin; vinblastine sulfate; vincristine sulfate;vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; and zorubicinhydrochloride.

Other anti-cancer drugs to be included within the methods orcompositions include, but are not limited to: 20-epi-1,25dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin;acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists;altretamine; ambamustine; amidox; amifostine; aminolevulinic acid;amrubicin; amsacrine; anagrelide; anastrozole; andrographolide;angiogenesis inhibitors; antagonist D; antagonist G; antarelix;anti-dorsalizing morphogenetic protein-1; antiandrogens, prostaticcarcinoma; antiestrogen; antineoplaston; antisense oligonucleotides;aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators;apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine;atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3;azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol;batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine;beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid;bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine;bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane;buthionine sulfoximine; calcipotriol; calphostin C; camptothecinderivatives; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docetaxel;docosanol; dolasetron; doxifluridine; doxorubicin; droloxifene;dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine;edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride;estramustine analogue; estrogen agonists; estrogen antagonists;etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine;fenretinide; filgrastim; finasteride; flavopiridol; flezelastine;fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex;formestane; fostriecin; fotemustine; gadolinium texaphyrin; galliumnitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine;glutathione inhibitors; hepsulfam; heregulin; hexamethylenebisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene;idramantone; ilmofosine; ilomastat; imatinib; imiquimod; immunostimulantpeptides; insulin-like growth factor-1 receptor inhibitor; interferonagonists; interferons; interleukins; iobenguane; iododoxorubicin;ipomeanol, 4-; iroplact; irsogladine; isobengazole; isohomohalicondrinB; itasetron; jasplakinolide; kahalalide F; lamellarin-N triacetate;lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin;letrozole; leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; loxoribine; lurtotecan; lutetiumtexaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A;marimastat; masoprocol; maspin; matrilysin inhibitors; matrixmetalloproteinase inhibitors; menogaril; merbarone; meterelin;methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine;mirimostim; mitoguazone; mitolactol; mitomycin analogues; mitonafide;mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene;molgramostim; cetuximab, human chorionic gonadotrophin; monophosphoryllipid A+mycobacterium cell wall sk; mopidamol; mustard anticancer agent;mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; nilutamide; nisamycin; nitric oxidemodulators; nitroxide antioxidant; nitrullyn; oblimersen;O⁶-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; paclitaxel protein-bound particles forinjectable suspension, albumin bound (ABRAXANE®); palauamine;palmitoylrhizoxin; pamidronic acid; panaxytriol; panomifene; parabactin;pazelliptine; pegaspargase; peldesine; pentosan polysulfate sodium;pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol;phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil;pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A; placetinB; plasminogen activator inhibitor; platinum complex; platinumcompounds; platinum-triamine complex; porfimer sodium; porfiromycin;prednisone; propyl bis-acridone; prostaglandin J2; proteasomeinhibitors; protein A-based immune modulator; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rohitukine;romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin;sarmustine; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine;senescence derived inhibitor 1; sense oligonucleotides; signaltransduction inhibitors; sizofiran; sobuzoxane; sodium borocaptate;sodium phenylacetate; solverol; somatomedin binding protein; sonermin;sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin1; squalamine; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; tallimustine; tamoxifen methiodide; tauromustine;tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomeraseinhibitors; temoporfin; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; translation inhibitors; tretinoin;triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron;turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors;ubenimex; urogenital sinus-derived growth inhibitory factor; urokinasereceptor antagonists; vapreotide; variolin B; velaresol; veramine;verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole;zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.

Specific second active agents particularly useful in the methods orcompositions include, but are not limited to, rituximab, oblimersen,infliximab, docetaxel, celecoxib, melphalan, dexamethasone, steroids,gemcitabine, cisplatinum, temozolomide, etoposide, cyclophosphamide,temodar, carboplatin, procarbazine, carmustine, tamoxifen, topotecan,methotrexate, gefitinib, paclitaxel, fluorouracil, leucovorin,irinotecan, capecitabine, interferon alpha, pegylated interferon alpha,cisplatin, thiotepa, fludarabine, carboplatin, liposomal daunorubicin,cytarabine, vinblastine, IL-2, GM-CSF, dacarbazine, vinorelbine,zoledronic acid, palmitronate, clarithormycin, busulphan, prednisone,bisphosphonate, arsenic trioxide, vincristine, doxorubicin, ganciclovir,estramustine sodium phosphate, clinoril, and etoposide.

Other specific second active agents particularly useful in the methodsor compositions include, but are not limited to, sorafenib, dabrafenib,vemurafenib, trametinib, cobimetinib, binimetinib, selumetinib,PD-325901, CI-1040 (PD184352), TAK-733, AT7867, AZD 8055, BX-912,silmitasertib, pictilisib, MK-2206, pilaralisib, gefitinib, erlotinib,lapatinib, osimertinib, CC-115, CC-223, OSI-027, AZD8055, sapanisertib,dactolisib, BGT226, voxtalisib, apitolisib, omipalisib, PF-04691502,gedatolisib, PP242, lenalidomide, pomalidomide, or CC-122.

Other specific second active agents particularly useful in the methodsor compositions include, but are not limited to, avelumab, durvalumab,MEDI0680, atezolizumab, BMS-936559, nivolumab, pembrolizumab,pidilizumab, PDR-001, sorafenib, cetuximab, panatumumab, erlotinib,trametinib, trastuzumab, CC-223, CC-122 or lapatinib.

In certain embodiments of the methods provided herein, use of a secondactive agent in combination with a solid form of Compound 1 describedherein may be modified or delayed during or shortly followingadministration of a solid form of Compound 1 described herein as deemedappropriate by the practitioner of skill in the art. In certainembodiments, subjects being administered a solid form of Compound 1described herein alone or in combination with other therapies mayreceive supportive care including antiemetics, myeloid growth factors,and transfusions of blood products, when appropriate. In someembodiments, subjects being administered a solid form of Compound 1described herein may be administered a growth factor as a second activeagent according to the judgment of the practitioner of skill in the art.

In certain embodiments, a solid form of Compound 1 described herein isadministered with gemcitabine, cisplatinum, 5-fluorouracil, mitomycin,methotrexate, vinblastine, doxorubicin, carboplatin, thiotepa,paclitaxel, paclitaxel protein-bound particles for injectablesuspension-albumin bound (ABRAXANE®), or docetaxel to patients withlocally advanced or metastatic urothelial carcinoma.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with a second active ingredient as follows:temozolomide to pediatric patients with relapsed or progressive braintumors or recurrent neuroblastoma; celecoxib, etoposide andcyclophosphamide for relapsed or progressive CNS cancer; temozolomide topatients with recurrent or progressive meningioma, malignant meningioma,hemangiopericytoma, multiple brain metastases, relapsed brain tumors, ornewly diagnosed glioblastoma multiforme; irinotecan to patients withrecurrent glioblastoma; carboplatin to pediatric patients with brainstem gliomas; procarbazine to pediatric patients with progressivemalignant gliomas; cyclophosphamide to patients with poor prognosismalignant brain tumors, newly diagnosed or recurrent glioblastomamultiforms; carmustine for high grade recurrent malignant gliomas;temozolomide and tamoxifen for anaplastic astrocytoma; or topotecan forgliomas, glioblastoma, anaplastic astrocytoma or anaplasticoligodendroglioma.

In certain embodiments, a solid form of Compound 1 described herein isadministered with methotrexate, cyclophosphamide, 5-fluorouracil,everolimus, paclitaxel, paclitaxel protein-bound particles forinjectable suspension-albumin bound (ABRAXANE®), lapatinib, trastuzumab,pamidronate disodium, eribulin mesylate, everolimus, gemcitabine,palbociclib, ixabepilone, ado-trastuzumab emtansine, pertuzumab,thiotepa, aromatase inhibitors, exemestane, selective estrogenmodulators, estrogen receptor antagonists, anthracyclines, emtansine,and/or pexidartinib to patients with metastatic breast cancer.

In certain embodiments, a solid form of Compound 1 described herein isadministered with temozolomide, doxorubicin, everolimus, fluorouracil,5-fluorouracil, or streptozocin to patients with neuroendocrine tumors.

In certain embodiments, a solid form of Compound 1 described herein isadministered with methotrexate, gemcitabine, cisplatin, cetuximab,5-fluorouracil, bleomycin, docetaxel or carboplatin to patients withrecurrent or metastatic head or neck cancer. In one embodiment, a solidform of Compound 1 as described herein provided herein is administeredwith cetuximab, to patients with head or neck cancer.

In certain embodiments, a solid form of Compound 1 described herein isadministered with gemcitabine, paclitaxel, paclitaxel protein-boundparticles for injectable suspension-albumin bound (ABRAXANE®),5-fluorouracil, everolimus, irinotecan, mitomycin C, sunitinib orerlotinib to patients with pancreatic cancer.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with colon cancer in combination withgetfitinib, erlotinib, oxaliplatin, 5-fluorouracil, irinotecan,capecitabine, cetuximab, ramucirumab, panitumumab, bevacizumab,leucovorin calcium, LONSURF, regorafenib, ziv-aflibercept, trametinib,paclitaxel, paclitaxel protein-bound particles for injectablesuspension-albumin bound (ABRAXANE®), and/or docetaxel. In certainembodiments, a solid form of Compound 1 as described herein providedherein is administered to patients with colon cancer in combination withbevacizumab, irinotecan hydrochloride, capecitabine, cetuximab,ramucirumab, oxaliplatin, cetuximab, fluorouracil, leucovorin calcium,trifluridine and tipiracil hydrochloride, panitumumab, regorafenib, orziv-aflibercept. In some embodiments, a solid form of Compound 1 asdescribed herein provided herein is administered to patients with coloncancer in combination with an EGFR inhibitor (for example cetuximab orerlotinib) and/or a BRAF inhibitor (for example, sorafenib, dabrafenib,or vemurafenib).

In certain embodiments, a solid form of Compound 1 described herein isadministered with capecitabine, cetuximab, erlotinib, trametinib, and/orvemurafenib to patients with refractory colorectal cancer or patientswho fail first line therapy or have poor performance in colon or rectaladenocarcinoma. In some embodiments, a solid form of Compound 1 asdescribed herein provided herein is administered to patients withrefractory colorectal cancer or patients who fail first line therapy orhave poor performance in colon or rectal adenocarcinoma in combinationwith an EGFR inhibitor (for example cetuximab or erlotinib) and a BRAFinhibitor (for example, sorafenib, dabrafenib, or vemurafenib). In someembodiments, a solid form of Compound 1 as described herein providedherein is administered to patients with refractory colorectal cancer orpatients who fail first line therapy or have poor performance in colonor rectal adenocarcinoma in combination with an anti-RSPO antibody.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with fluorouracil, leucovorin, trametiniband/or irinotecan to patients with Stage Ma to IV colorectal cancer orto patients who have been previously treated for metastatic colorectalcancer. In some embodiments, a solid form of Compound 1 as describedherein provided herein is administered to patients with Stage Ma to IVcolorectal cancer or to patients who have been previously treated formetastatic colorectal cancer, in combination with an EGFR inhibitor (forexample cetuximab or erlotinib) and a BRAF inhibitor (for example,sorafenib, dabrafenib, or vemurafenib). In certain embodiments, a solidform of Compound 1 as described herein provided herein is administeredto patients with refractory colorectal cancer in combination withcapecitabine, xeloda, trametinib, oxaliplatin and/or irinotecan. In someembodiments, a solid form of Compound 1 as described herein providedherein is administered to patients with refractory colorectal cancer, incombination with an EGFR inhibitor (for example cetuximab or erlotinib)and a BRAF inhibitor (for example, sorafenib, dabrafenib, orvemurafenib). In certain embodiments, a solid form of Compound 1 asdescribed herein provided herein is administered with capecitabine,trametinib, and/or irinotecan to patients with refractory colorectalcancer or to patients with unresectable or metastatic colorectalcarcinoma. In some embodiments, a solid form of Compound 1 as describedherein provided herein is administered to patients with refractorycolorectal cancer or to patients with unresectable or metastaticcolorectal carcinoma, in combination with an EGFR inhibitor (for examplecetuximab or erlotinib) and a BRAF inhibitor (for example, sorafenib,dabrafenib, or vemurafenib).

In certain embodiments, a solid form of Compound 1 described herein isadministered alone or in combination with interferon alpha,5-fluorouracil/leucovorin or capecitabine to patients with unresectableor metastatic hepatocellular carcinoma; or with cisplatin and thiotepa,or with sorafenib to patients with primary or metastatic liver cancer.In certain embodiments, a solid form of Compound 1 as described hereinprovided herein is administered alone or in combination with sorafenib,sunitinib, erlotinib, and/or sirolimus, to patients with unresectable ormetastatic hepatocellular carcinoma; or with sorafenib, sunitinib,erlotinib, and/or rapamycin to patients with primary or metastatic livercancer. In some embodiments, a solid form of Compound 1 as describedherein provided herein is administered to patients with primary,unresectable, or metastatic liver cancer, in combination with an immunecheckpoint inhibitor (for example, an anti-CTLA4, anti-PD1, anti-PD-L1,anti-Tim-3, or anti-Lag-3 antibody) or a BRAF inhibitor (for example,sorafenib, dabrafenib, or vemurafenib). In some such embodiments, theanti-PD-1 or anti-PD-L1 antibody is avelumab, durvalumab, MEDI0680,atezolizumab, BMS-936559, nivolumab, pembrolizumab, pidilizumab, orPDR-001. In certain embodiments, a solid form of Compound 1 as describedherein provided herein is administered alone or in combination withlenalidomide, pomalidomide or CC-122 to patients with primary,unresectable or metastatic hepatocellular carcinoma. In certainembodiments, a solid form of Compound 1 as described herein providedherein is administered alone or in combination CC-223 to patients withprimary, unresectable or metastatic hepatocellular carcinoma.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with cisplatin/5-fluorouracil, ramucirumab,docetaxel, doxorubicin hydrochloride, fluorouracil injection,trastuzumab, and/or mitomycin C to patients with gastric (stomach)cancer.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with an immune checkpoint inhibitor (forexample, an anti-CTLA4, anti-PD1, anti-PD-L1, anti-Tim-3, or anti-Lag-3antibody) and/or a BRAF inhibitor (for example, sorafenib, dabrafenib,or vemurafenib) to patients with various types or stages of melanoma. Insome embodiments, a solid form of Compound 1 as described hereinprovided herein is administered in combination with aldesleukin,cobimetinib, dabrafenib, dacarbazine, IL-2, talimogene laherparepvec,recombinant interferon alfa-2b, ipilimumab, pembrolizumab, lapatinib,trametinib, nivolumab, peginterferon alfa-2b, aldesleukin, dabrafenib,and/or vemurafenib to patients with various types or stages of melanoma.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with doxorubicin, paclitaxel, paclitaxelprotein-bound particles for injectable suspension-albumin bound(ABRAXANE®), vinblastine or pegylated interferon alpha to patients withKaposi's sarcoma.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with methotrexate, mechlorethaminehydrochloride, afatinib dimaleate, pemetrexed, bevacizumab, carboplatin,cisplatin, ceritinib, crizotinib, ramucirumab, pembrolizumab, docetaxel,vinorelbine tartrate, gemcitabine, paclitaxel, paclitaxel protein-boundparticles for injectable suspension-albumin bound (ABRAXANE®),erlotinib, geftinib, and/or irinotecan to patients with non-small celllung cancer.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with carboplatin and irinotecan to patientswith non-small cell lung cancer.

In certain embodiments, a solid form of Compound 1 described herein isadministered with docetaxel to patients with non-small cell lung cancerwho have been previously treated with carboplatin/etoposide andradiotherapy.

In certain embodiments, a solid form of Compound 1 described herein isprovided herein is administered in combination with carboplatin and/ordocetaxel, or in combination with carboplatin, pacilitaxel, paclitaxelprotein-bound particles for injectable suspension-albumin bound(ABRAXANE®), and/or thoracic radiotherapy to patients with non-smallcell lung cancer.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with docetaxel to patients with stage IIIBor IV non-small cell lung cancer.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with oblimersen, methotrexate,mechlorethamine hydrochloride, etoposide, topotecan or doxorubicin topatients with small cell lung cancer.

In certain embodiments, a solid form of Compound 1 described herein anddoxetaxol are administered to patients with small cell lung cancer whowere previously treated with carbo/VP 16 and radiotherapy.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with various types or stages of ovarian cancersuch as peritoneal carcinoma, papillary serous carcinoma, refractoryovarian cancer or recurrent ovarian cancer, in combination withcarboplatin, doxorubicin, gemcitabine, cisplatin, capecitabine,paclitaxel, paclitaxel protein-bound particles for injectablesuspension-albumin bound (ABRAXANE®), dexamethasone, avastin,cyclophosphamide, topotecan, olaparib, thiotepa, or a combinationthereof.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with various types or stages of prostatecancer, in combination with capecitabine, 5-fluorouracil plusleucovorin, gemcitabine, irinotecan plus gemcitabine, cyclophosphamide,vincristine, dexamethasone, GM-CSF, celecoxib, ganciclovir, paclitaxel,paclitaxel protein-bound particles for injectable suspension-albuminbound (ABRAXANE®), docetaxel, estramustine, denderon, abiraterone,bicalutamide, cabazitaxel, degarelix, enzalutamide, goserelin,leuprolide acetate, mitoxantrone hydrochloride, prednisone,sipuleucel-T, radium 223 dichloride, or a combination thereof.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with various types or stages of renal cellcancer, in combination with capecitabine, IFN, tamoxifen, IL-2, GM-CSF,celecoxib, or a combination thereof.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with various types or stages of gynecologic,uterus or soft tissue sarcoma cancers in combination with IFN,dactinomycin, doxorubicin, imatinib mesylate, pazopanib, hydrochloride,trabectedin, a COX-2 inhibitor such as celecoxib, and/or sulindac.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with various types or stages of solid tumors incombination with celecoxib, etoposide, cyclophosphamide, docetaxel,apecitabine, IFN, tamoxifen, IL-2, GM-CSF, or a combination thereof.

In certain embodiments, a solid form of Compound 1 described herein isadministered alone or in combination with vinorelbine to patients withmalignant mesothelioma, or stage IIIB non-small cell lung cancer withpleural implants or malignant mesothelioma syndrome.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with A navitoclax, venetoclax and/orobatoclax to patients with lymphoma and other blood cancers.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with arsenic trioxide, fludarabine,carboplatin, daunorubicin, cyclophosphamide, cytarabine, doxorubicin,idarubicin, mitoxantrone hydrochloride, thioguanine, vincristine, and/ortopotecan to patients with acute myeloid leukemia, including refractoryor relapsed or high-risk acute myeloid leukemia.

In certain embodiments, a solid form of Compound 1 described herein isadministered in combination with liposomal daunorubicin, topotecanand/or cytarabine to patients with unfavorable karotype acutemyeloblastic leukemia.

In certain embodiments, a solid form of Compound 1 described herein isadministered alone or in combination with a second active ingredientsuch as vinblastine or fludarabine, chlorambucil, bleomycin, brentuximabvedotin, carmustine, chlorambucil, cyclophosphamide, dacarbazine,doxorubicin, lomustine, mechlorethamine hydrochloride, prednisone,procarbazine hydrochloride or vincristine to patients with various typesof lymphoma, including, but not limited to, Hodgkin's lymphoma,non-Hodgkin's lymphoma, cutaneous T-Cell lymphoma, cutaneous B-Celllymphoma, diffuse large B-Cell lymphoma or relapsed or refractory lowgrade follicular lymphoma.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with various types or stages of multiplemyeloma in combination with dexamethasone, zoledronic acid, pamitronate,GM-CSF, clarithromycin, vinblastine, melphalan, busulphan,cyclophosphamide, IFN, prednisone, bisphosphonate, celecoxib, arsenictrioxide, peginterferon alfa-2b, vincristine, carmustine, bortezomib,carfilzomib, doxorubicin, panobinostat, lenalidomide, pomalidomide,thalidomide, plerixafor or a combination thereof.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with various types or stages of multiplemyeloma in combination with chimeric antigen receptor (CAR) T-cells.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with relapsed or refractory multiple myeloma incombination with doxorubicin, vincristine and/or dexamethasone.

In certain embodiments, a solid form of Compound 1 described herein isadministered to patients with scleroderma or cutaneous vasculitis incombination with celecoxib, etoposide, cyclophosphamide, docetaxel,capecitabine, IFN, tamoxifen, IL-2, GM-CSF, or a combination thereof.

Also encompassed herein is a method of increasing the dosage of ananti-cancer drug or agent that can be safely and effectivelyadministered to a patient, which comprises administering to the patient(e.g., a human) a solid form of Compound 1 described herein. Patientsthat can benefit by this method are those likely to suffer from anadverse effect associated with anti-cancer drugs for treating a specificcancer of the skin, subcutaneous tissue, lymph nodes, brain, lung,liver, bone, intestine, colon, heart, pancreas, adrenal, kidney,prostate, breast, colorectal, or combinations thereof. Theadministration of a solid form of Compound 1 described herein alleviatesor reduces adverse effects which are of such severity that it wouldotherwise limit the amount of anti-cancer drug.

In one embodiment, a solid form of Compound 1 described herein isadministered daily in an amount ranging from about 0.1 to about 150 mg,from about 1 to about 100 mg, from about 2 to about 50 mg, or from about1 to about 10 mg prior to, during, or after the occurrence of theadverse effect associated with the administration of an anti-cancer drugto a patient. In certain embodiments, a solid form of Compound 1described herein is administered in combination with specific agentssuch as heparin, aspirin, coumadin, anti-Factor Xa, or G-CSF to avoidadverse effects that are associated with anti-cancer drugs such as butnot limited to thromboembolism, neutropenia or thrombocytopenia.

In one embodiment, a solid form of Compound 1 described herein isadministered to patients with diseases and disorders associated with orcharacterized by, undesired angiogenesis in combination with additionalactive ingredients, including, but not limited to, anti-cancer drugs,anti-inflammatories, antihistamines, antibiotics, and steroids.

In another embodiment, encompassed herein is a method of treating,preventing and/or managing cancer, which comprises administering a solidform of Compound 1 described herein in conjunction with (e.g. before,during, or after) conventional therapy including, but not limited to,surgery, immunotherapy, biological therapy, radiation therapy, or othernon-drug based therapy presently used to treat, prevent or managecancer. The combined use of the compound provided herein andconventional therapy may provide a unique treatment regimen that isunexpectedly effective in certain patients. Without being limited bytheory, it is believed that a solid form of Compound 1 described hereinmay provide additive or synergistic effects when given concurrently withconventional therapy.

As discussed elsewhere herein, encompassed herein is a method ofreducing, treating and/or preventing adverse or undesired effectsassociated with conventional therapy including, but not limited to,surgery, chemotherapy, radiation therapy, hormonal therapy, biologicaltherapy and immunotherapy. A solid form of Compound 1 as provided hereinand other active ingredient can be administered to a patient prior to,during, or after the occurrence of the adverse effect associated withconventional therapy.

Cycling Therapy

In certain embodiments, the prophylactic or therapeutic agents providedherein are cyclically administered to a patient. Cycling therapyinvolves the administration of an active agent for a period of time,followed by a rest for a period of time, and repeating this sequentialadministration. Cycling therapy can reduce the development of resistanceto one or more of the therapies, avoid, or reduce the side effects ofone of the therapies, and/or improves the efficacy of the treatment.

Consequently, in certain embodiments, a solid form of Compound 1provided herein is administered daily in a single or divided dose in afour to six week cycle with a rest period of about a week or two weeks.In certain embodiments, a solid form of Compound 1 provided herein isadministered daily in a single or divided doses for one to tenconsecutive days of a 28 day cycle, then a rest period with noadministration for rest of the 28 day cycle. The cycling method furtherallows the frequency, number, and length of dosing cycles to beincreased. Thus, encompassed herein in certain embodiments is theadministration of a solid form of Compound 1 provided herein for morecycles than are typical when it is administered alone. In certainembodiments, a solid form of Compound 1 provided herein is administeredfor a greater number of cycles that would typically cause dose-limitingtoxicity in a patient to whom a second active ingredient is not alsobeing administered.

In one embodiment, a solid form of Compound 1 provided herein isadministered daily and continuously for three or four weeks at a dose offrom about 0.1 to about 150 mg/day followed by a break of one or twoweeks.

In another embodiment, a solid form of Compound 1 provided herein isadministered intravenously and a second active ingredient isadministered orally, with administration of a solid form of Compound 1described herein occurring 30 to 60 minutes prior to a second activeingredient, during a cycle of four to six weeks. In certain embodiments,the combination of a solid form of Compound 1 provided herein and asecond active ingredient is administered by intravenous infusion overabout 90 minutes every cycle. In certain embodiments, one cyclecomprises the administration from about 0.1 to about 150 mg/day of asolid form of Compound 1 provided herein and from about 50 to about 200mg/m²/day of a second active ingredient daily for three to four weeksand then one or two weeks of rest. In certain embodiments, the number ofcycles during which the combinatorial treatment is administered to apatient is ranging from about one to about 24 cycles, from about two toabout 16 cycles, or from about four to about three cycles.

Pharmaceutical Compositions and Routes of Administration

Solid forms of Compound 1 described herein can be administered to asubject orally, topically or parenterally in the conventional form ofpreparations, such as capsules, microcapsules, tablets, granules,powder, troches, pills, suppositories, injections, suspensions, syrups,patches, creams, lotions, ointments, gels, sprays, solutions andemulsions. Suitable formulations can be prepared by methods commonlyemployed using conventional, organic or inorganic additives, such as anexcipient (e.g., sucrose, starch, mannitol, sorbitol, lactose, glucose,cellulose, talc, calcium phosphate or calcium carbonate), a binder(e.g., cellulose, methylcellulose, hydroxymethylcellulose,polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic,polyethyleneglycol, sucrose or starch), a disintegrator (e.g., starch,carboxymethylcellulose, hydroxypropylstarch, low substitutedhydroxypropylcellulose, sodium bicarbonate, calcium phosphate or calciumcitrate), a lubricant (e.g., magnesium stearate, light anhydrous silicicacid, talc or sodium lauryl sulfate), a flavoring agent (e.g., citricacid, menthol, glycine or orange powder), a preservative (e.g., sodiumbenzoate, sodium bisulfate, methylparaben or propylparaben), astabilizer (e.g., citric acid, sodium citrate or acetic acid), asuspending agent (e.g., methylcellulose, polyvinyl pyrrolidone oraluminum stearate), a dispersing agent (e.g.,hydroxypropylmethylcellulose), a diluent (e.g., water), and base wax(e.g., cocoa butter, white petrolatum or polyethylene glycol). Theeffective amount of the solid forms of Compound 1 described herein inthe pharmaceutical composition may be at a level that will exercise thedesired effect; for example, about 0.005 mg/kg of a subject's bodyweight to about 10 mg/kg of a subject's body weight in unit dosage forboth oral and parenteral administration.

The dose of a solid form of Compound 1 to be administered to a subjectis rather widely variable and can be subject to the judgment of ahealth-care practitioner. In general, the solid forms of Compound 1 canbe administered one to four times a day in a dose of about 0.005 mg/kgof a subject's body weight to about 10 mg/kg of a subject's body weightin a subject, but the above dosage may be properly varied depending onthe age, body weight and medical condition of the subject and the typeof administration. In one embodiment, the dose is about 0.01 mg/kg of asubject's body weight to about 10 mg/kg of a subject's body weight,about 0.1 mg/kg of a subject's body weight to about 10 mg/kg of asubject's body weight, about 1 mg/kg of a subject's body weight to about10 mg/kg of a subject's body weight or about 1 mg/kg of a subject's bodyweight to about 5 mg/kg of a subject's body weight. In one embodiment,one dose is given per day. In any given case, the amount of the solidform of Compound 1 administered will depend on such factors as thesolubility of the active component, the formulation used and the routeof administration. In one embodiment, application of a topicalconcentration provides intracellular exposures or concentrations ofabout 0.01-10 M.

In another embodiment, provided herein are methods for the treatment orprevention of a disease or disorder comprising the administration ofabout 1 mg/day to about 1000 mg/day, about 1 mg/day to about 750 mg/day,about 1 mg/day to about 500 mg/day, about 1 mg/day to about 250 mg/dayor about 100 mg/day to about 1000 mg/day of a solid form of Compound 1described herein to a subject in need thereof.

In another embodiment, provided herein are unit dosage formulations thatcomprise between about 1 mg and 1000 mg, about 5 mg and about 1000 mg,about 10 mg and about 1000 mg, about 25 mg and about 1000 mg, about 50mg and about 1000 mg, about 100 mg and about 1000 mg, or about 250 mgand about 1000 mg of a solid form of Compound 1 described herein.

A solid forms of Compound 1 described herein can be administered once,twice, three, four or more times daily. In a particular embodiment,doses of 600 mg or less are administered as a once daily dose and dosesof more than 600 mg are administered twice daily in an amount equal toone half of the total daily dose.

In another embodiment, provided herein are unit dosage formulations thatcomprise between about 1 mg and 200 mg, about 35 mg and about 1400 mg,about 125 mg and about 1000 mg, about 250 mg and about 1000 mg, or about500 mg and about 1000 mg of a solid form of Compound 1 described herein.

In a particular embodiment, provided herein are unit dosage formulationscomprising about 100 mg or 400 mg of a solid form of Compound 1described herein.

In another embodiment, provided herein are unit dosage formulations thatcomprise 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 30 mg, 35 mg, 50 mg, 70 mg,100 mg, 125 mg, 140 mg, 175 mg, 200 mg, 250 mg, 280 mg, 350 mg, 500 mg,560 mg, 700 mg, 750 mg, 1000 mg or 1400 mg of a solid form of Compound 1described herein.

The solid forms of Compound 1 described herein can be administered once,twice, three, four or more times daily. In a particular embodiment,doses of 600 mg or less are administered as a once daily dose and dosesof more than 600 mg are administered twice daily in an amount equal toone half of the total daily dose.

The solid forms of Compound 1 described herein can be administeredorally for reasons of convenience. In one embodiment, when administeredorally, a solid form of Compound 1 is administered with a meal andwater. In another embodiment, the solid form of Compound 1 is dispersedin water or juice (e.g., apple juice or orange juice) and administeredorally as a suspension.

The solid forms of Compound 1 described herein can also be administeredintradermally, intramuscularly, intraperitoneally, percutaneously,intravenously, subcutaneously, intranasally, epidurally, sublingually,intracerebrally, intravaginally, transdermally, rectally, mucosally, byinhalation, or topically to the ears, nose, eyes, or skin. The mode ofadministration is left to the discretion of the health-carepractitioner, and can depend in part upon the site of the medicalcondition.

In one embodiment, provided herein are capsules containing a solid formof Compound 1 described herein without an additional carrier, excipientor vehicle.

In another embodiment, provided herein are compositions comprising aneffective amount of a solid form of Compound 1 described herein and apharmaceutically acceptable carrier or vehicle, wherein apharmaceutically acceptable carrier or vehicle can comprise anexcipient, diluent, or a mixture thereof. In one embodiment, thecomposition is a pharmaceutical composition.

The compositions can be in the form of tablets, chewable tablets,capsules, solutions, parenteral solutions, troches, suppositories andsuspensions and the like. Compositions can be formulated to contain adaily dose, or a convenient fraction of a daily dose, in a dosage unit,which may be a single tablet or capsule or convenient volume of aliquid. In one embodiment, the solutions are prepared from water-solublesalts, such as the hydrochloride salt. In general, all of thecompositions are prepared according to known methods in pharmaceuticalchemistry. Capsules can be prepared by mixing a solid form of Compound 1described herein with a suitable carrier or diluent and filling theproper amount of the mixture in capsules. The usual carriers anddiluents include, but are not limited to, inert powdered substances suchas starch of many different kinds, powdered cellulose, especiallycrystalline and microcrystalline cellulose, sugars such as fructose,mannitol and sucrose, grain flours and similar edible powders.

Tablets can be prepared by direct compression, by wet granulation, or bydry granulation. Compression of the solid forms of Compound 1 describedherein may not reduce or modulate the activity of the administered drugto a patient. Their formulations usually incorporate diluents, binders,lubricants and disintegrators as well as the compound. Typical diluentsinclude, for example, various types of starch, lactose, mannitol,kaolin, calcium phosphate or sulfate, inorganic salts such as sodiumchloride and powdered sugar. Powdered cellulose derivatives are alsouseful. Typical tablet binders are substances such as starch, gelatinand sugars such as lactose, fructose, glucose and the like. Natural andsynthetic gums are also convenient, including acacia, alginates,methylcellulose, polyvinylpyrrolidine and the like. Polyethylene glycol,ethylcellulose and waxes can also serve as binders.

A lubricant might be necessary in a tablet formulation to prevent thetablet and punches from sticking in the die. The lubricant can be chosenfrom such slippery solids as talc, magnesium and calcium stearate,stearic acid and hydrogenated vegetable oils. Tablet disintegrators aresubstances that swell when wetted to break up the tablet and release thecompound. They include starches, clays, celluloses, algins and gums.More particularly, corn and potato starches, methylcellulose, agar,bentonite, wood cellulose, powdered natural sponge, cation-exchangeresins, alginic acid, guar gum, citrus pulp and carboxymethyl cellulose,for example, can be used as well as sodium lauryl sulfate. Tablets canbe coated with sugar as a flavor and sealant, or with film-formingprotecting agents to modify the dissolution properties of the tablet.The compositions can also be formulated as chewable tablets, forexample, by using substances such as mannitol in the formulation.

When it is desired to administer a solid form of Compound 1 describedherein as a suppository, typical bases can be used. Cocoa butter is atraditional suppository base, which can be modified by addition of waxesto raise its melting point slightly. Water-miscible suppository basescomprising, particularly, polyethylene glycols of various molecularweights are in wide use.

The effect of the solid form of Compound 1 described herein can bedelayed or prolonged by proper formulation. For example, a slowlysoluble pellet of the solid form of Compound 1 described herein can beprepared and incorporated in a tablet or capsule, or as a slow-releaseimplantable device. The technique also includes making pellets ofseveral different dissolution rates and filling capsules with a mixtureof the pellets. Tablets or capsules can be coated with a film thatresists dissolution for a predictable period of time. Even theparenteral preparations can be made long-acting, by dissolving orsuspending the solid form of Compound 1 described herein in oily oremulsified vehicles that allow it to disperse slowly in the serum.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form A, including substantially pure Form A.

In certain embodiments, the pharmaceutical compositions provided hereincomprise HCl Salt Form 1, including substantially pure starting materialHCl Salt Form.

In certain embodiments, the pharmaceutical compositions provided hereincomprise HCl Salt Form 1, including substantially pure HCl Salt Form 1.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form B, including substantially pure Form B.

In certain embodiments, the pharmaceutical compositions provided hereincomprise HCl Salt Form 2, including substantially pure HCl Salt Form 2.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form C, including substantially pure Form C.

In certain embodiments, the pharmaceutical compositions provided hereincomprise HCl Salt Form 3, including substantially pure HCl Salt Form 3.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form D, including substantially pure Form D.

In certain embodiments, the pharmaceutical compositions provided hereincomprise HCl Salt Form 4, including substantially pure HCl Salt Form 4.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form E, including substantially pure Form E.

In certain embodiments, the pharmaceutical compositions provided hereincomprise HCl Salt Form 5, including substantially pure HCl Salt Form 5.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form F, including substantially pure Form F.

In certain embodiments, the pharmaceutical compositions provided hereincomprise HCl Salt Form 6, including substantially pure HCl Salt Form 6.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form G, including substantially pure Form G.

In certain embodiments, the pharmaceutical compositions provided hereincomprise HCl Salt Form 7, including substantially pure HCl Salt Form 7.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form H, including substantially pure Form H.

In certain embodiments, the pharmaceutical compositions provided hereincomprise HCl Salt Form 8, including substantially pure HCl Salt Form 8.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form I, including substantially pure Form I.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form Y, including substantially pure Form Y.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form Z, including substantially pure Form Z.

In certain embodiments, the pharmaceutical compositions provided hereincomprise an amorphous solid, e.g. free base, HCl salt, citrate salt, orother salt described herein, including the substantially pure amorphoussolid.

In certain embodiments, the pharmaceutical compositions provided hereincomprise a mixture of one or more solid form(s) of Compound 1, includingForm A, Form B, Form C, Form D, Form E, Form F, Form G, Form H, Form I,Form Y, Form Z, HCl Salt Form 1, HCl Salt Form 2, HCl Salt Form 3, HClSalt Form 4, HCl Salt Form 5, HCl Salt Form 6, HCl Salt Form 7, HCl SaltForm 8 or an amorphous solid described herein, wherein every possiblecombination of the solid forms of Compound 1 is possible.

Examples

The following Examples are presented by way of illustration, notlimitation. The following abbreviations are used in descriptions andexamples:

ACN: Acetonitrile Am: Amorphous

AmPhos: p-Dimethylamino phenylditbutylphosphine

API: Active Pharmaceutical Ingredient

Boc: tert-Butoxycarbonyln-BuOH: n-Butanoldba: Dibenzylidene acetoneDBU: 1,8-Diazabicyclo[5.4.0]undec-7-ene

DCM: Dichloromethane DIPEA: N,N-Diisopropylethylamine DMAc:N,N-Dimethylacetamide DMF: N,N-Dimethylformide DMSO: Dimethylsulfoxide

DSC: Differential Scanning calorimetry

DVS: Dynamic Vapor Sorption

EDTA: Ethylenediamine tetraacetateESI: Electrospray ionizationEtOAc: Ethyl acetate

EtOH: Ethanol FTIR: Fourier Transform Infrared Spectroscopy

HPLC: High performance liquid chromatography

IPA: 2-Propanol

IPAc: Isopropyl acetateLCMS: Liquid Chromatography with Mass Spectroscopy

MeCN Acetonitrile

MEK: Methyl ethyl ketone

MeOH: Methanol

2-MeTHF: 2-Methyl tetrahydrofuranmp: Melting pointMS: Mass spectrometryMTBE: tert-Butyl methyl ether

NB S: N-Bromosuccinimide

NMP: N-Methyl-2-pyrrolidoneNMR: Nuclear magnetic resonance

RH: Relative Humidity RT: Room Temperature Rx Recrystallization S:Solvent SDTA: Single Differential Thermal Analysis

SM: Starting materialS-SegPhos (S)-(−)-5,5-Bis(diphenylphosphino)-4,4-bi-1,3-benzodioxole

TA: Thermal Analysis

Tf: Triflate or trifluoromethanesulfonylTFA: Trifluoroacetic acid

TFE: 2,2,2-Trifluoroethanol TGA: Thermogravimetric Analysis

TGA-MS/TG-MS: Thermogravimetric Analysis coupled with Mass Spectroscopy

THF: Tetrahydrofuran

TLC: Thin layer chromatography

XRPD: X-Ray Powder Diffraction Synthetic Examples

The following non-limiting synthetic examples show methods for thepreparation of Compound 1. ACD/NAME (Advanced Chemistry Development,Inc., Ontario, Canada) and/or Chemdraw (Cambridgesoft, Perkin Elmer,Waltham, Mass.) was used to generate names for chemical structures andChemdraw was used to draw the chemical structures.

In one embodiment, Compound 1 is synthesized in a manner as described inExample 53 of U.S. Pat. No. 9,512,124, which is hereby incorporated byreference in its entirety.

Compound 1 Salt Screening and Selection

Compound 1 salt form screening was conducted using small volumeapproaches. The pKa of Compound 1 is 5.14. Several counter ions werechosen for salt formation including glycolic, malic, citric, tartaric,phosphoric, maleic, benenesulfonic, methansulonic, toluenesulfonic,sulfuric, hydrochloric acids with various solvents.

Free base Compound 1 is hydrate material (a monohydrate). TGA weightloss amounted to 2.9% weight loss prior to decomposition, and DSC showedtwo endothermic peaks, broad at low temperature due to dehydration andthen melting peak at 182° C. The crystal form remained unchanged aftereither slurry in water. The free base is stable in solution (pH 1.2 to7.5) at 40° C. It has chemical and physical stability in solid stateunder stress conditions up to seven weeks. Under dry conditions, thehydrate form changed to partial or hemihydrates. The salt form likelyimproves the solid state properties and the pH-dependent solubility. Thecrystal form of monohydrate remained unchanged unless dried (<5% RH) ormaintained at higher temperature (>60° C.). The monohydrate free base isslightly hygroscopicity.

The solid samples were examined using X-ray diffractometer (SmartLab,Rigaku). The detector was equipped with a photomultiplier withpreamplifier X-ray detection technology. The samples were scanned from 3to 40° 2θ, at a step size 0.02° 2θ and a time per step of 20 seconds.The tube voltage and current were 40 KV and 44 mA, respectively. Thesample was transferred from sample container onto zero backgroundXRD-holder and gently ground.

TGA analyses were carried out on a TA Instruments TGA Q5000.Approximately 1.50 mg of samples was placed in a tared platinum oraluminum pan, automatically weighed, and inserted into the TGA furnace.The samples were heated at a rate of 10° C./min, to final temperature of300° C. The purge gas was nitrogen for balance at ca. 10 cc/min and forfurnace at ca 90 cc/min, respectively.

DSC analyses were conducted on a TA Instruments Q2000. The calibrationstandard was indium. A sample 1.50 mg in weight was placed into a taredTA DSC pan, and weight accurately recorded. Crimped pans were used foranalysis and the samples were heated under nitrogen (50 cc/min) at arate of 10° C./min, up to a final temperature of 300° C. The data wereprocessed using a thermal analyzer (Universal Analyzer 2000, TAInstruments).

Proton NMR was used to study the chemical shifts of compound resultedfrom salt formation. Proton NMR was performed using Bruker Advance 300Ultrashield™ equipped with automated sample (B-ACS 60). Dimethylsulfoxide-d₆ (DMSO-d₆) was used as a solvent for NMR analysis.Acquisition time was about 16 seconds,

Dynamic vapor sorption (DVS) was measured using DVS advantage (SurfaceMeasurement Systems Ltd). The samples were tested under isotherm (25°C.) at a targeted RH of 0 to 95% full cycle in step mode. For anisotherm test, the chamber temperature was maintained by a water bath atconstant 25.0±1.0° C. The relative humidity in the sample chamber wasgenerated by combining different flows of wet and dry nitrogen withvariable flow rates. The analysis was performed in 10% RH increments.Sampling rate was 1 sec save data rate is 20 sec. The dm/dt (%) valuewas set at 0.001 with a dm/dt window of 5 min., a minimum stabilityduration time of 10 min, and a maximum stage time of 180 min. Thesample's equilibrium weight corresponding to each RH was recorded. Asorption isotherm was obtained by plotting equilibrium moisture contentversus RH.

1.00 gram of Compound 1 free base was dissolved in 10 mL of methanol.100 μL of the stock solution was then added into each well on 96-wellplate. Acid solutions were added with molar 1:1 ratio into each well onto plate, one acid to 8 wells in the same row. After drying of theplate, aliquots of 400 μL of 8 different solvents were added into wellonto the plate in column fashion. The plates were then covered andallowed to evaporate in an operating laboratory fume hood under ambientconditions of temperature and humidity. Solvents were used for thescreening including ethanol, 2-propanol, 3-methyl-butanol, acetonitrile,methyl tert-butyl ether (MTBE), acetone, water, ethyl acetate.

The starting non-salt form of Compound 1 free base was characterized byXRPD, TGA, and DSC. It is crystalline monohydrate and here designed tobe Form 1.

Powder X-ray diffraction was performed on Compound 1, and the profile isshown in FIG. 89.

In FIG. 90, the TGA thermogram of Compound 1 shows that about a 2.9%weight loss was observed at relative low temperature (<75° C.) due todehydration. The final weight loss is from decomposition of the drugcompound.

In FIG. 91, the DSC thermogram of Compound 1 showed that the crystallinesolid has a broad endothermic event at relative low temperaturecorresponding to dehydration/desolvation, and the endothermic peak withonset and peak temperature of 174.6 and 182.1° C., respectively, withenthalpy of 52.0 J/g due to the melt of the dehydrated form.

The profiles of DVS showed the sample (Compound 1) is slightlyhygroscopic (<4.3%) from 0-95% RH with respect to monohydrate free baseas shown in FIG. 92. The monohydrate free base converted to anhydrouswhen first placed through a drying cycle at zero or very low humidity.Then ˜2.6% (wt) of water was gained when the solid particles wereexposed to increasing relative humidity up to 25% RH, converted backmonohydrate in this range of humidity. Additional ˜1.7% moisturesorption is slowly and steadily gained from 25 to 95% RH. Duringdesorption cycle from 95 down to 5% RH, the loss of water content isvery slowly 1.5%, the monohydrate structure maintained. Then, theremaining ˜2.8% wt of water was suddenly released from the sample asrelative humidity decreased from 5% RH to dry. The level of 3.0% watercontent is corresponding to monohydrate.

The adsorption/desorption are almost reversible above 30% RH. Below 30%RH, the release of water during desorption is more difficult than uptake during sorption.

Proton NMR of Compound 1 was examined in DMSO and shown in FIG. 6.Compound 1 free base (1.00 gram) was dissolved in methanol (10.0 mL).Aliquots of 100 μL of the solution were then distributed into each wellonto a 96-well plate (1.0 mL flat bottom clear glass inserts). Elevenacids including glycolic, malic, citric, tartaric, phosphoric, maleic,benenesulfonic, methansulonic, toluenesulfonic, sulfuric, hydrochloricacids were added with molar ratio of 1:1 into wells, see Table 1.Solvent was evaporated in an operation laboratory fume hood underambient conditions of temperature and humidity. After dryness, thesolvents for crystallization were introduced, see Table 1. Then, theplate was covered with a round-welled-cap mat w/silicone/PTFE liner toallow slow evaporation and crystallization at ambient environment.

HCl was initially found during the solubility study of Compound 1freebase in simulated gastric fluid (SGF) solution. About 30 mg ofCompound 1 free base was weighed into a glass vial, and 1 mL of SGF wasintroduced. The mixture soon became clear solution. Overnight,precipitation occurred. The solid particles were collected viafiltration and were characterized. The XRPD profile is different fromthe freebase, as shown in FIG. 94. TGA showed about 3.3% weight loss atrelatively low temperature (<70° C.) prior to decomposition (FIG. 95).The DSC profile had two endothermic events 1) at relatively lowtemperature due to dehydration and melting dehydrated form with onsetand peak temperatures of 209.3 and 230.5° C., respectively (FIG. 96).Dynamic vapor sorption was performed on this sample at isotherm and isshown in FIG. 97. ¹H NMR showed that chemical shifts were observed onhydrogen in purine and benzene ring, suggested the salt formation (FIG.98).

Dynamic vapor sorption (DVS) showed the HCl salt is low hygroscopicity(<1.0%) from 0-95% RH with respect to monohydrate HCl as shown in FIG.97. The HCl monohydrate converted to anhydrous when first placed througha drying cycle at zero or very low humidity. Then ˜2.9% (wt) of waterwas gained when the solid particles were exposed to increasing relativehumidity up to 25% RH, converted back monohydrate in this range ofhumidity. Additional ˜1.7% moisture sorption was slowly and steadilygained from 25 to 95% RH. During desorption cycle from 95 down to 5% RH,the loss of water content was very slowly ˜1.5%, the monohydratestructure maintained. Then, the remaining ˜2.8% wt of water was rapidlyreleased from the sample as relative humidity decreased from 5% RH todry. The level of 3.0% water content is corresponding to monohydrate.

The adsorption/desorption were almost reversible above 30% RH. Below 30%RH, the release of water during desorption was more difficult than uptake during sorption. Hysteresis was observed between the sorption anddesorption curves below 25% RH.

Sample from well# H2 containing both Compound 1 free base and sulfuricacid was analyzed by ¹H NMR in DMSO-d₆, FIG. 99. ¹H NMR showed thatchemical shifts were observed on hydrogen in purine and benzene ring,suggested the salt formation. Selected samples were analyzed by TGA andDSC, FIG. 100 to FIG. 105. TGA profiles all showed initial weight losses(2.5-3.2%) at relatively low temperature, however, were not samebehaviors. DSC profiles from these wells also showed broad endothermicevents at relatively low temperatures, but were not same profile.Sulfate salts are solvates, depending on the use of solvent forcrystallization.

Crystalline mesylate salts from SVSS in various solvents were found andXRPD profiles of crystalline citrate salts from various solvents arevery similar. A representative XRPD profile of crystalline mesylatesalts from SVSS in EtOAC is shown in FIG. 106. Sample containing bothCompound 1 free base and methansulfonic acid was analyzed by ¹H NMR inDMSO-_(d6), FIG. 107. ¹H NMR showed that chemical shifts were observedon hydrogen in purine and benzene ring, suggested the salt formation.Selected samples were analyzed by TGA and DSC, FIG. 108 to FIG. 117. TGAprofiles f all showed initial weight losses (1.4-2.2%) at relatively lowtemperature, and slightly different behaviors. DSC profiles from thesewells also showed broad endothermic events at relatively lowtemperatures, but different profiles after desolvation.

Crystalline citrate salts from SVSS in various solvents were found andXRPD profiles of crystalline citrate salts from SVSS are shown in FIG.118 to FIG. 125. XRPD profiles from ethanol (A9), IPA (B9),3-methyl-2-butanol (C9), acetonitrile (D9), MTBE (E9), and acetone (F9)appear similar (citrate form 1), as shown in FIG. 126 and FIG. 127. XRPDprofile from SVSS in EtOAc (H9) was different (FIG. 128, citrate form2). XRPD profile from SVSS in water (FIG. 124) was very close to thosein FIG. 126. Sample containing both Compound 1 free base and citric acidwas analyzed by ¹H NMR in DMSO-d₆, FIG. 129. ¹H NMR showed that nochemical shifts were observed in hydrogen in purine and benzene ring,suggested that salt formation is weak interaction in solution phase andcan be regarded as co-crystal. Selected samples were analyzed by TGA andDSC, FIG. 130 to FIG. 139. TGA profiles all showed minimum initialweight losses (<0.5%) at relatively low temperature prior todecomposition. DSC profiles from these wells also showed a singleendothermic due to melt with onset and peak temperatures of ˜203 and˜211° C., respectively.

A sample containing both Compound 1 free base and phosphoric acid wasanalyzed by ¹H NMR in DMSO-d₆, FIG. 140. ¹H NMR showed that no chemicalshifts were observed in hydrogen in purine and benzene ring suggestedthat salt formation may not likely in solution phase. Different crystalstructures were found from SVSS, suggested that co-crystals of phosphatewith Compound 1 were formed.

The X-ray powder diffraction patterns of wells B4 and B6 were similar,designated as phosphate Form 1A FIG. 141.

X-ray powder diffraction of wells B7 and B10 were different and alsodifferent from the Form 1A, designated as Form 1B and 1C, respectively,as shown in FIG. 142.

The HCl salt monohydrate (1): 240 mg Compound 1 was weighed into a 4-mLglass vial, and then 4.60 mL of 0.1N HCl in water was introduced. Themixture became clear. The solution was filtered via a 0.22 μm filter andthe supernatant was placed under hood for crystallization. Soon,precipitation occurred. The solid was collected via filtration.

The solid sample was analyzed to be monohydrate (1), and the XRPDprofile is similar with the one from the solubility study of free basein SGF but has better crystallinity, as mL glass vial and then 3.10 mLof 0.5N HCl in water was added. The mixture became clear. Additional 5.0mL of water was added. The solution was filtered via a 0.22 μm filter.The supernatant was placed under hood for crystallization. Soon,precipitation occurred. The solid was collected via filtration.

303.7 mg Compound 1 was weighed in a 20 mL glass vial and then 10 mL ofSGF was added. The mixture became clear. Solid particles of Compound 1HCl salt were added into the vial, as seeds. The suspension was keptagitation on LabQuake rotation for 24 hours. Solid particles werecollected via filtration.

The anhydrous form was not observed in solution precipitation process.The dehydration was performed on XRD-DSC stage.

170 mg Compound 1 was weighed into a 4 mL glass vial and then 3.3 mL of0.1M H₂SO₄ in EtOAc was introduced. The mixture became gummy/gellingmaterial immediately. After drying, the solid was collected and analyzedby)(RFD, TGA and DSC, as shown in FIG. 153 to FIG. 155.

105 mg Compound 1 was weighed into a 4 mL glass vial and 2.0 mL of 0.1MH₂SO₄ in water was introduced. The mixture became gel-like material, andaddition of 1 mL of water was added. The material was still oil-likesticky.

138 mg Compound 1 was weighed into a 4 mL glass vial and then 1.0 mL ofEtOAc was added to dissolve the material first. Then, 2.60 mL of 0.1Mmethanesulfonic acid in EtOAc was introduced, and precipitate appearedimmediately. The solid was collected via filtration and dried at 40° C.under vacuum overnight, and then analyzed by)(RFD, TGA and DSC, as shownin FIG. 156 to FIG. 158.

34 mg Compound 1 was weighed into a 4-mL glass vial and then 0.65 mL of0.1M methanesulfonic acid in acetonitrile was added. No clear solutionwas achieved, however, new solid phase was obviously observed. The solidwas collected via filtration and dried at 40° C. under vacuum overnight,and then analyzed by)(RFD, TGA and DSC, as shown in FIG. 159 to FIG.161.

Mesylate salt form: After slurry in water, the XRPD profile is slightlydifferent as shown in FIG. 162.

Meslyate salt was also studied under moisture using dynamic vapoursorption (DVS). After sorption and desorption cycle (FIG. 163), the XRPDpattern is slightly different from the starting material, as shown inFIG. 164.

114 mg Compound 1 was weighed in a glass vial and then 0.6 mL EtOAcsolvent was added. The mixture became clear solution after agitation.2.20 mL of 0.1N citric in water was added into the solution, andcotton-like precipitates appeared immediately. The solid was collectedvia filtration and characterized, as shown in FIG. 165 to FIG. 167. TGAshowed little weight loss (<0.2%) at relatively low temperature prior todecomposition. DSC showed a single endothermic peak due to melting withonset and peak temperatures of 205.2 and 209.5° C., respectively, withenthalpy of 240.7 J/g.

95 mg Compound 1 was weighed in a glass and then 0.5 mL acetone solventwas added to dissolve the material. Then 1.8 mL of 0.1N citric inacetone was added. The clear solution was placed under hood forcrystallization. Soon, precipitates appeared.

The solid was collected via filtration and characterized, as shown inFIG. 168 to FIG. 170. XRPD profile was similar to form 1. TGA showedlittle weight loss (<0.4%) at relatively low temperature prior todecomposition. DSC showed a single endothermic peak due to melting withonset and peak temperatures of 205.3 and 209.5° C., respectively, withenthalpy of 250.2 J/g.

208 mg Compound 1 was weighed in a glass and then 1.0 mL acetone solventwas added to dissolve the material. Then 4.0 mL of 0.1N citric in waterwas added. The clear solution was placed under hood for crystallization.Soon, precipitates appeared. The solid was collected via filtration andcharacterized. TGA showed little weight loss (<0.4%) at relatively lowtemperature prior to decomposition. DSC showed a single endothermic peakdue to melting with onset and peak temperatures of 206.0 and 211.4° C.,respectively, with enthalpy of 257.7 J/g.

TGA showed nearly no weight losses prior to decomposition, however, bothNMR and GC analysis showed the presence of 4000-5000 ppm acetone.

43.02 mg Compound 1 was weighed in a glass and then 1.0 mL Ethanolsolvent was added to dissolve the material (not completely). Then 0.82mL of 0.1N citric in water was added and the mixture became clear. Theclear solution was placed under hood for crystallization. Soon,precipitates appeared. Additional 1.0 mL of water added. The solid wascollected via filtration and characterized, as shown in FIG. 174 to FIG.176.

XRPD profile was different from forms 1, 2, and 3. TGA showed littleweight loss (<0.3%) at relatively low temperature prior todecomposition. DSC showed a single endothermic peak due to melting withonset and peak temperatures of 211.2 and 214.8° C., respectively, withenthalpy of 277.1 J/g.

45.74 mg Compound 1 was weighed in a glass and then 1.0 mL IPA solventwas added to dissolve the material (cloudy). Then 0.87 mL of 0.1N citricin water was added, and the mixture became clear. The clear solution wasplaced under hood for crystallization. Shortly, precipitates appeared.

The solid was collected via filtration and characterized, TGA showedlittle weight loss (<0.3%) at relatively low temperature prior todecomposition. However, DSC showed a broad endothermic peak atrelatively low temperature due possibly to desolvation and melting peakwith onset and peak temperatures of 207.9 and 212.7° C., respectively,with enthalpy of 190.7 J/g.

51.4 mg Compound 1 was weighed in a glass and then 1.0 mL of 0.1N citricacid in water was added. The suspension was kept agitation at ambientfor conversion and crystallization. (Addition 1 mL water). The solid wascollected via filtration and characterized, as shown in FIG. 180 to FIG.182. TGA showed little weight loss (<0.1%) at relatively low temperatureprior to decomposition. DSC showed a single of melting peak with onsetand peak temperatures of 204.2 and 207.6° C., respectively, withenthalpy of 249.9 J/g.

Citrate salt showed low hygroscopicity as demonstrated from dynamicvapor sorption (DVS) study (FIG. 183). From SVSS screening, the solidforms with citric acid in various solvents resulted in similar XRPDprofile, which are likely an isostructural solvates. TGA showed slightweight loss (<0.5%) at relatively low temperature, DSC showed singleendothermic peak with onset and peak temperatures of 205.3 and 209.5° C.Citrate salt produced in several solvents including ethanol, IPA,acetone, and EtOAc.

The solubility of HCl hydrate Form (2), citrate salt Form Z (2) and freebase was determined in water, simulated gastric fluid (SGF), simulatedintestinal fluid (SIF), and 0.5% HPMC in 0.25% Tween 80. The solubilityin water varied, depending on pH. It can be seen from Table 3 that HClsalt monohydrate has highest solubility in water (sparingly soluble inwater) at pH 3.65. The solubility of citrate salt and free base in waterare 0.252 and 0.003 mg/mL, respectively, depending on pH. The pH inwater media was determined by both counter-ions and solubility. HClsalts resulted in lowest pH 3.65 in water while citrate resulted in pHrelative high pH=4.61. Solubility of HCl salt form in SGF has effect ofcommon ions; however, free base has significant high kinetic solubilityin SGF, followed by citrate salt. Solubility of these salts as well asfree base in SIF is quite low, practically insoluble in SIF.

TABLE 3 Solubility of various salt forms in water, SGF, SIF and 0.5%HPMC in 0.25 Tween80 Conc. at Conc. at 2 Hrs 24 Hrs pH at Forms Media(mg/mL) (mg/mL) final Free Base Water 0.000 0.003 8.11 SGF >31 3.4801.93 SIF 0.000 0.002 7.33 0.5% HPMC/0.25% Tween80 0.208 0.178 4.23 HClSalt Water 1.664 1.726 3.65 SGF 0.361 0.351 1.17 SIF 0.000 0.000 7.220.5% HPMC/0.25% Tween80 3.878 4.696 3.06 Citrate Water 0.076 0.252 4.61SGF 1.954 1.691 1.44 SIF 0.000 0.000 7.32 0.5% HPMC/0.25% Tween80 0.1860.325 3.81

The solubility of HCl and citrate salt was also determined inbiorelevant media in comparison with free base. In the presence ofsurfactants (Sodium taurocholate and Lecithin), the solubility values ofHCl salt, citrate and free base are similar in both FeSSIF and FaSSIF(Table 4).

TABLE 4 Solubility of HCl and citrate salts in biorelevant media incomparison with free base. Free Base HCl Salt Citrate Conc. Conc. Conc.Media mg/mL pH mg/mL pH mg/mL pH FeSSIF 1.920 4.95 1.978 4.85 1.957 4.71FaSSIF 0.025 6.37 0.043 6.12 0.024 4.65 FaSSGF 3.847 2.77 0.121 1.820.227 1.93 FeSSGF 0.002 6.23 0.001 5.66 0.001 5.21

Among the free base, HCl and citrate salt, there did not no significantdifference in the solid state properties. They are all chemically andphysically stable in solid state. The data herein suggest that thesolubility of free base, HCl and citrate salt depends on the pH, theeffect of common ions, and the presence of surfactants. No conclusive PKresults were obtained from single dog PK comparison study. All of solidforms showed similar dissolution profiles in both FeSSIF and FaSSIF,(FIG. 186 and FIG. 187) and demonstrated similar dissolution profiles ofBIC in both 0.01N HCl and 0.001N HCl.

TABLE 5 Solid state stability of salt forms and free base Chemical (40°C./ Chemical Physical 75% RH) (60° C.) (40° C./ Physical Solid Forms RRT= 2.2 RRT2.2 75% RH) (60° C.) Free Base, — <0.03% Stable Dehydration(−), hydrate reversible + RH HCl, hydrate2 0.02% 0.05% Stable StableCitrate 0.02% 0.06% Stable Stable

Solid Forms Analytical Methods—Free Base

A polymorph screen of Compound 1 was performed to investigate whetherdifferent solid forms could be generated under various conditions, suchas different solvents, temperature and humidity changes.

The solvents used in the polymorph screen were either HPLC or reagentgrade, including acetonitrile (MeCN), MeCN/water (1:1), n-butanol(n-BuOH), absolute ethanol (EtOH), ethanol/water (1:1), methanol (MeOH),2-propanol (IPA), ethyl acetate (EtOAc), methyl acetate (MeOAc),dichloromethane (DCM), methyl ethyl ketone (MEK), methyl t-butyl ether(MTBE), heptane, toluene, methyl acetate (MeOAc), isopropyl acetate(IPAc), methyl isobutyl ketone (MIBK), 2-methyltetrahydrofuran(2-MeTHF), 1,4-dioxane, tetrahydrofuran (THF), THF/water (1:1), andwater.

A weighed sample of Compound 1 was treated with a known volume of a testsolvent. The resulting mixture was agitated for about 1 day at roomtemperature. If all of the solids appeared to be dissolved by visualinspection, the estimated solubility was calculated based on the totalvolume of solvent used to give a complete solution. If solids werepresent, a known volume of filtrate was evaporated to dryness and theweight of the residue was measured to estimate the solubility.

All of the solid samples generated in the polymorph screen were analyzedby XRPD. XRPD analysis was conducted on a PANalytical Empyrean X-raypowder diffractometer using Cu Kα radiation at 1.54 Å.

The PANalytical Empyrean instrument was equipped with a fine focus X-raytube. The voltage and amperage of the X-ray generator were set at 45 kVand 40 mA, respectively. The divergence slits were set at 1/16° and ⅛°,and the receiving slit was set at 1/16°. Diffracted radiation wasmeasured using a Pixel 2D detector. A theta-two theta continuous scanwas set at step size 0.013 from 3° to 40° 2θ with sample spinning rateat 4. A sintered alumina standard was used to check the peak positions.

DSC analyses were performed on a TA Discovery Differential Scanningcalorimeter. Indium was used as the calibration standard. Approximately1-5 mg of sample was placed into a DSC pan. The sample was heated undernitrogen at a rate of 10° C./min, up to a final temperature of 220° C.Melting points were reported as the extrapolated onset temperatures.

TGA analyses were performed on a TA Discovery ThermogravimetricAnalyzer. Approximately 2-10 mg of accurately weighed sample was placedon a pan and loaded into the TGA furnace. The sample was heated undernitrogen at a rate of 10° C./min, up to a final temperature of 220° C.

Morphology analysis of the samples was carried out on an Evex Mini-SEM.Small amounts of samples were dispersed on a sample holder, coated withgold using an Evex Mini Au Sputter Coater, and imaged with 300× to 1000×magnification.

Hygroscopicity was determined on a Surface Measurement Systems DVS. Asample size of 5-20 mg was loaded into the DVS instrument sample pan andthe sample was analyzed on a DVS automated sorption analyzer at roomtemperature. The relative humidity was increased from 0% to 90% RH at10% RH step, then decreased in a similar manner to accomplish a fulladsorption/desorption cycle.

¹H NMR spectra were obtained on a Bruker 300 MHz NMR spectrometer.Samples were dissolved in DMSO-D⁶ and analyzed with 8-64 scans.

Karl Fischer (KF) water content was measured using a Metrohm KFcoulometric oven titrator equipped with an oven sample processor. Theoven temperature was set as 100° C.

Equilibration/Slurry and Evaporation Experiments

Equilibrium and evaporation experiments carried out at room temperature.If solids were present after 1 day, they were filtered using a 0.45 μmPTFE filter and air-dried before analysis. The remaining supernatant wasevaporated to dryness and the solids were isolated for analysis.

Equilibration and evaporation experiments at 50° C. were carried out byadding an excess of solid Compound 1 to up to 1 mL of a test solvent.The resulting mixture was agitated for 1 day at room temperature and 1day at 50° C. separately. Upon reaching equilibrium, the saturatedsupernatant solution was removed, filtered using 0.45 μm PTFE filtersand allowed to evaporate in an open vial under nitrogen at roomtemperature and 50° C., respectively. The solid resulting from theequilibration was isolated and air-dried before analysis.

TABLE 6 Summary Equilibrium (EQ) and Evaporation (EV) Results Solvent EQat RT EV at RT EQ at 50° C. EV at 50° C. 1,4-dioxane — amorphous —amorphous 2-MeTHF — amorphous — amorphous acetone — amorphous —amorphous DCM — amorphous — amorphous MeCN A + C — C + A — MeCN/water(1:1) A + C — C + A — EtOAc — amorphous H + A amorphous EtOH — amorphousH + A amorphous EtOH/water (1:1) A + H — E — water A — A — Heptane A — A— IPA D amorphous F F IPAc — amorphous — amorphous MEK — amorphous —amorphous MeOAc — amorphous — amorphous MeOH — amorphous — amorphousMIBK — amorphous — amorphous MTBE — amorphous G — n-BuOH — — — amorphousTHE — amorphous — amorphous THF/water (1:1) — amorphous — amorphousToluene A — * amorphous+ * — not analyzable *not enough crystallinematerial for accurate characterization

Anti-Solvent Recrystallization and Cooling Recrystallization Experiments

For cooling recrystallization, each of the selected solvents wassaturated with solid Compound 1 at 65° C. The solvents included MeCN,MeCN/water (1:1), EtOH, EtOH/water (1:1), IPA, and THF/water (1:1). Thesolution was stirred for about 10 minutes, filtered using a 0.45 μm PTFEsyringe filter, and then cooled to about −15° C. by placing the vialsinto a freezer. The solid resulting from the recrystallization wasisolated and air-dried before analysis. For cooling recrystallization,each of the selected solvents (MeOH, EtOH, and EtOH/water) was saturatedwith Compound 1 at 60° C. The solution was stirred at 60° C. for 10minutes, filtered using a 0.45 μm PTFE syringe filter, and then cooledto room temperature naturally and then placed into a refrigerator. Thesolid resulting from the recrystallization was isolated and air-driedbefore analysis.

TABLE 7 Cooling Recrystallization Results Solvent Conditions Form byXRPD MeCN 65° C. to −15° C. I MeCN/water (1:1) 65° C. to −15° C. C EtOH65° C. to −15° C. — EtOH/water (1:1) 65° C. to −15° C. — THF/water (1:1)65° C. to −15° C. — IPA 65° C. to −15° C. —

no precipitation/not enough material for analysis

For anti-solvent recrystallization, the selected solvents MeCN and MeOHwere saturated with solid Compound 1 at the room temperature. Once thesolid was completely dissolved, a portion of the solution was filteredinto a vial containing a selected anti-solvent (water). The mixture wascooled to 4° C. by placing the vials into a refrigerator. The solidresulting from the recrystallization was isolated and air-dried beforeanalysis. For anti-solvent recrystallization, the selected solvents(MeOH, EtOH, IPA, and EtOAc) were saturated with Compound 1 at 60° C.Once the solid was completely dissolved, a portion of the solution wasfiltered into a pre-heated vial and a selected anti-solvent (water,MTBE, or heptane) was added at 60° C. The mixture was cooling to roomtemperature naturally and then placed into a refrigerator. The solidresulting from the recrystallization was isolated and air-dried beforeanalysis.

TABLE 8 Experiments to Generate Materials for Characterization Form bySolvent Experimental Conditions XRPD none starting with Form A, dried invacuum B oven at 40° C. MeCN Slurry starting with Form A at 50° C. C IPASlurry starting with Form A at RT D EtOH/water (1:1) Slurry startingwith Form A at 50° C. H IPA Slurry starting with Form A at 50° C. F MTBESlurry starting with Form A at 50° C. G EtOH Slurry starting with Form Aat 50° C. H MeCN Recrystallization from saturated solution I of Form Aat 65° C. cooled to −15° C.

MeOH, EtOH, EtOH/water, IPA, and EtOAc were used as single or primarysolvents. Water, MTBE, and heptanes were used as anti-solvent. Theresults are summarized in Table 6. Only crystallizations using water asanti-solvents generated Form A. All other solvents or solventcombinations afforded similar solvate forms as observed duringequilibration experiment.

TABLE 9 Anti-solvent Recrystallization Results Solvent Anti-solventRatio Form by XRPD MeCN water 1:10 C + A MeOH water 1:2  amorphous

Summary of Polymorphic Forms

A total of nine crystalline forms and an amorphous form for Compound 1as a free base were found during this polymorph screen study. The stackplot of XRPD patterns for the nine crystalline forms are shown in FIG.1, and the physical characteristics are summarized in Table 10. The XRPDpattern of the amorphous form is shown in FIG. 40.

TABLE 10 Summary of Solid Forms and Amorphous Form for Compound 1 FreeBase Representative DSC peaks TGA Form Description Conditions (° C.) (%wt loss) DVS or other notes A mono- Starting material 117, 182 2.8 4.7wt % water hydrate uptake up to 90% RH B anhydrate Drying Form A at 40°C. 182 <1.3 Converts to Form A or drying Form C at at >20% RH 50-60° C.in vacuum oven C MeCN solvate EQ in MeCN 165, 186 6.6 Converts to Form Aat >20% RH D IPA solvate EQ at RT in IPA 154, 185 7.4 — E solvate or EQat 50° C. in 104, 115, 13.7 — hydrate EtOH/water (1:1) 119, 165 F IPAsolvate EQ or EV at 50° C. in 153 14.3 — IPA G MTBE solvate EQ at 50° C.in MTBE 148, 161 8.7 — H solvate or EQ at 50° C. in 163, 187 6.5Converts to Form A hydrate EtOH/water (1:1), at >20% RH EtOH, EtOAc IMeCN Cooling crystallization 75, 183 2.3 — solvate in MeCN amorphous EVfrom most — — 6.9 wt % water solvents at RT and uptake up to 50° C. 90%RH —: not applicable or not available EQ: equilibration EV: evaporation

Form A

Approximate solubility of free base Form A in various solvents atambient temperature was estimated as described above. The results aresummarized in Table 11. Free base Form A was found to be most soluble(>100 mg/mL) in acetone, EtOAc, MeOAc, and THF. Form A was very soluble(>50 mg/mL) in 1,4-dioxane, 2-MeTHF, DCM, MeCN/water (1:1), IPAc, MEK,MeOH, MIBK, and THF/water (1:1). Form A showed some solubility (>20mg/mL) in EtOH, MTBE, n-BuOH, (>10 mg/mL) in IPA, Toluene, (>3 mg/mL) inMeCN, and EtOH/water (1:1). Form A showed low solubility (<1 mg/mL) inwater and heptane.

TABLE 11 Approximately Solubility of Compound 1 Free Base Form A at RoomTemperature Approximate Solubility Solvent (mg/mL) 1,4-dioxane >502-MeTHF >50 acetone >100 DCM >50 MeCN 3 MeCN/water (1:1) >50 EtOAc >100EtOH 25 EtOH/water (1:1) 5 water <1 Heptane <1 IPA 18 IPAc >50 MEK >50MeOAc >100 MeOH >50 MIBK >50 MTBE 34 n-BuOH 25 THE >100 THF/water(1:1) >50 Toluene 17

Equilibrium experiments at 50° C. resulted in Form A in water andheptane. A unique form designated Form E was obtained from Form A inEtOH/water (1:1). A unique form designated Form F was obtained from FormA in IPA. A unique form designated Form G was obtained from Form A inMTBE. A mixture of Form A and Form C was obtained in MeCN and MeCN/water(1:1). A mixture of Form A and Form H was obtained in EtOAc and EtOH.Form F was also obtained from Form A from the evaporation at 50° C. fromIPA. Evaporation in toluene resulted in a mixture of the amorphous andlow crystalline material (unknown form). All other evaporationexperiments at 50° C. resulted in the amorphous form of Compound 1.

Cooling recrystallization experiments were performed as described above.The solvents included MeCN, MeCN/water (1:1), EtOH, EtOH/water (1:1),THF/water (1:1), and IPA. The results are summarized in Table 7. Thesolids obtained from MeCN/water (1:1) were confirmed to be Form C. Thesolids obtained from MeCN were confirmed to be a unique form designatedForm I. The remaining solvents did not precipitate after 14 days at −15°C.

Recrystallizations with anti-solvents were performed as described above.MeCN and MeOH were used as the primary solvent. Water was used asanti-solvents. The results are summarized in Table 10. Using XRPD, thesolids obtained from MeCN/water were confirmed to be a mixture of Form Cand Form A. The solids obtained from MeOH/water were confirmed to beamorphous.

Form A is a monohydrate. This form was mostly obtained fromrecrystallization or slurry experiments in aqueous or “water-rich”solvent systems.

Form A can also be obtained by conversion from Form B, Form C, and FormH by exposure to ambient conditions having greater than about 20%relative humidity (RH).

Form A converts to the anhydrous Form B upon drying at below 10% RH orat elevated temperature.

Form A has a crystalline XRPD pattern as shown in FIG. 2. TGA and DSCthermograms of Form A are shown in FIG. 4 and FIG. 5, respectively. TheDSC thermogram showed two events with a first having an onsettemperature of about 94° C. and a maximum of about 117° C., attributedto dehydration and a second having an onset temperature of about 174° C.and maximum of about 182° C., corresponding to melt/decomposition. TGAweight loss of 2.8% was observed up to melt. The ¹H NMR spectrum of FormA was consistent with Compound 1 structure with no significantdegradation or residual solvent (see FIG. 7).

The moisture sorption/desorption behavior of Form A was determined byDVS. The results are summarized in FIG. 6A-FIG. 6B. A steep weightchange over 3% was observed between 10 and 30% RH. A similar weightchange was observed between 10 to 0% RH upon desorption, which isconsistent with a hydrate. Additional water uptake of approximately 1.4wt % was observed between 30-90% RH, suggesting the hydrate is slightlyhygroscopic.

FIG. 1 provides an XRPD pattern of Form A. A list of X-Ray DiffractionPeaks for Form A is provided below in Table 12.

TABLE 12 X-Ray Diffraction Peaks for Form A Relative Two-theta angle (°)d Space (Å) Intensity (%) 3.20123 27.60013 7.39 7.332486 12.05637 64.828.513228 10.38668 72.85 10.74741 8.23198 22.68 11.05949 8.00038 5.1312.67103 6.98627 8.99 12.97577 6.82287 9.21 13.43725 6.58957 13.4713.77375 6.42933 11.89 14.45398 6.12825 30.43 14.69756 6.02723 36.7215.94064 5.55991 25.02 16.88512 5.25098 22.32 17.07588 5.19274 31.1617.32333 5.11912 12.45 17.72045 5.00529 10.31 18.18509 4.87844 10018.65178 4.75741 11.04 20.29581 4.37561 8.64 20.73897 4.2831 19.8221.0281 4.22485 21.03 21.26496 4.17833 38.12 22.11363 4.01985 19.3422.66107 3.92397 12.51 22.87923 3.88704 10.28 23.1537 3.84158 13.1923.61127 3.76816 3.67 24.00033 3.70795 6.78 24.83366 3.58538 7.1425.53212 3.48886 4.65 26.1405 3.40903 5.74 26.40372 3.37564 12.726.79629 3.32707 14.69 27.86908 3.20139 6.64 28.08561 3.1772 13.7528.84895 3.09484 21.58 29.42683 3.03286 4.2 29.77657 3.00051 7.18 31.4442.8451 10.84 31.80169 2.81158 2.81 32.56458 2.74971 2.03 33.145532.70283 1.97 33.60055 2.66727 2.6 33.93018 2.63992 2.51 34.21827 2.620525.43 34.66795 2.58755 6.41 36.13494 2.4858 2.98 36.4681 2.46385 337.24883 2.41398 2.35 37.73477 2.38401 1.03 38.93764 2.31308 3.1839.50587 2.28112 1.35

FIG. 3 is an SEM image of Form A.

Form B

Form B was obtained from drying Form A at about 40° C. under vacuum.Form B can also be obtained from drying Form C at 50-60° C. undervacuum. Form B converts to Form A at ambient conditions that includegreater than about 20% RH. Form B had a crystalline XRPD pattern asshown in FIG. 8. TGA and DSC thermograms of Form B obtained from acetoneare shown in FIG. 9 and FIG. 10, respectively. The TGA weight loss of0.1 wt % corresponded to one DSC peak around with an onset of about 174°C. and maximum of about 182° C. and corresponded to themelt/decomposition. These observations suggested that Form B is ananhydrate of Compound 1.

A list of X-Ray Diffraction Peaks for Form B is provided below in Table13.

TABLE 13 X-Ray Diffraction Peaks for Form B Relative Two-theta angle (°)d Space (Å) Intensity (%) 6.890295 12.82908 19.62 8.730049 10.1292 21.510.47572 8.44486 10.12 11.62559 7.61205 23.62 12.00448 7.37264 21.8513.5532 6.53345 14.74 13.79915 6.41755 42.51 14.0533 6.30206 23.3514.22065 6.22827 12.68 16.2888 5.44184 5.17 16.91908 5.24051 5 17.525575.0605 20.97 18.04876 4.91497 24.18 18.44801 4.8095 8.46 19.144474.63607 15.55 19.4722 4.55878 100 19.98866 4.44214 68.99 20.762194.27836 30.81 21.07678 4.21521 11.17 22.10397 4.02159 4.97 22.680523.92065 10.77 23.33598 3.81199 6.7 25.15811 3.53695 2.82 26.026453.42371 18.19 26.71736 3.33672 5.74 27.3612 3.25965 6.19 28.394363.14335 6.28 28.82505 3.09479 3.02 29.19153 3.0593 4.72 30.11261 2.967798.81 30.95864 2.88859 7.7 31.51091 2.83921 7.06 31.8305 2.81143 6.03

Form C

Form C was obtained from equilibration of Form A in MeCN or MeCN/waterat room temperature or 50° C. Form C is also obtainable from process asolution of Compound 1 in MeTHF. MeTHF (10 vol) was distilled undervacuum at constant volume with addition of MeCN (˜20 vol) to removeMeTHF (230 torr/46° C.). At the end no more than 5 vol % MeTHF was inthe batch. The solids crystallized during the distillation. The batchwas cooled, aged, filtered, and dried under vacuum at no higher than 30°C. Form C had a crystalline XRPD pattern as shown in FIG. 11. TGA andDSC thermograms of Form C obtained from MeCN/water are shown in FIG. 12and FIG. 13, respectively. The TGA weight loss of 6.6 wt % correspondedto broad DSC peak around 165° C. and can be attributed to desolvation inForm C. The DSC peak with onset temperature of 180° C. and a maximum ofabout 186° C. corresponded to the melt/decomposition. The ¹H-NMRspectrum was obtained for the Form C sample and was consistent withstructure though with high amount of MeCN present (FIG. 14). Thetheoretical MeCN content of a mono-solvate of Compound 1 is 6.7 wt %,matching the TGA weight loss observed. These observations suggested thatForm C is an acetonitrile mono-solvate of Compound 1. Form C in ambienttemperatures of greater than about 20% RH resulted conversion to Form A.

A list of X-Ray Diffraction Peaks for Form C is provided below in

Table 14.

TABLE 14 X-Ray Diffraction Peaks for Form C Relative Two-theta angle (°)d Space (Å) Intensity (%) 3.146478 28.08028 4.17 7.733216 11.4325 76.488.895852 9.94078 39.19 10.33009 8.56359 100 13.28054 6.66697 3.8213.65962 6.48279 15.14 14.15782 6.25577 2.18 14.55961 6.08403 7.7314.80553 5.98352 3.64 15.01642 5.89995 5.47 15.30885 5.78791 20.2215.50492 5.71515 23.7 15.66686 5.65644 26.8 16.94314 5.23313 17.5317.35927 5.10861 32.43 17.79869 4.98346 16.72 18.28813 4.85118 77.4218.73191 4.73724 5.29 19.48359 4.55614 4.1 19.94116 4.45262 27.3420.72705 4.28553 4.27 21.1145 4.20776 6.33 21.40394 4.15151 22.8722.09642 4.02295 1.75 22.43447 3.96309 5.19 22.65575 3.92488 5.5723.14226 3.84346 8.41 23.91671 3.72073 4.29 24.59886 3.61907 3.0225.02821 3.55795 33.9 25.51834 3.49072 5.57 25.81841 3.45082 7.7526.14386 3.4086 34 26.72919 3.33527 15.86 26.94224 3.30664 2.91 27.179083.27836 2.5 27.6642 3.22463 14 28.47546 3.13199 21.49 29.3905 3.036533.38 29.79318 2.99639 1.72 30.33114 2.94446 5.58 30.90051 2.8915 5.2631.31067 2.85455 6.86 32.44982 2.75689 16.95 32.96986 2.71458 5.5633.58429 2.66631 4.06 34.27597 2.61407 1.27 35.41687 2.53243 1.4235.94476 2.49644 2.65 36.24054 2.47674 3.82 37.12225 2.41992 1.0937.89885 2.3721 1.29 38.90024 2.31331 2.87

Form D

Form D was obtained from recrystallization equilibration of Form A inIPA at room temperature. Form D had a crystalline XRPD pattern as shownin FIG. 15. TGA and DSC thermograms of Form D are shown in FIG. 17 andFIG. 18, respectively. The TGA weight loss of approximately 7.4 wt %corresponded to a broad DSC peak around 154° C. and can be attributed toloss of solvent in Form D. The smaller DSC peak with onset temperatureof 175° C. and maximum of about 185° C. corresponded to themelt/decomposition. The ¹H-NMR spectrum was obtained for the Form Dsample and was consistent with structure and contained IPA (see FIG.19). These observations suggested that Form D is most likely an IPAsolvate of Compound 1.

A list of X-Ray Diffraction Peaks for Form D is provided below in Table15.

TABLE 15 X-Ray Diffraction Peaks for Form D Relative Two-theta angle (°)d Space (Å) Intensity (%) 3.143947 28.10288 5.24 5.896986 14.98764 18.387.358024 12.01459 45.8 8.731096 10.12798 41.92 10.13588 8.72723 96.4811.11409 7.96121 16.27 13.65702 6.48402 8.38 14.78355 5.99237 25.5615.11394 5.86211 31.94 16.34076 5.42466 6.65 16.61001 5.33732 11.1317.56252 5.04994 29.87 18.06277 4.9112 100 19.22765 4.61621 12.8119.77163 4.49041 11.36 20.38698 4.35624 34.4 21.48209 4.13658 28.8822.14167 4.01483 13.22 22.34634 3.97852 12.64 23.95561 3.71477 12.9624.27345 3.66381 7.94 24.97168 3.56588 10.23 26.20157 3.40122 16.126.87895 3.31703 6.79 27.3269 3.26366 9.21 27.60494 3.22875 8.8328.19356 3.16528 7.43 28.6337 3.11762 8.48 30.89808 2.89411 8 31.445522.84497 5.69 32.89163 2.72312 1.93 33.58744 2.66828 3.68 34.6305 2.590262.32 37.15969 2.41957 2.88 34.25 2.6183 1.6 35.39 2.5363 0.6 35.872.5034 2.8 36.55 2.4588 1.5 36.81 2.4415 2.7 37.06 2.4261 2.1 37.772.3820 2.8 38.60 2.3323 1.8

Form E

Form E was obtained from equilibration of Form A in EtOH/water (1:1) at50° C. Form E had a crystalline XRPD pattern as shown in FIG. 20. TGAand DSC thermograms of Form E are shown in FIG. 21 and FIG. 22,respectively. The TGA weight loss of 13.7 wt % corresponded to smallbroad DSC peak around 104° C. and can be attributed to loss of solventin Form E. These observations suggested that Form E is a solvate orhydrate containing ethanol.

A list of X-Ray Diffraction Peaks for Form E is provided below in Table16.

TABLE 16 X-Ray Diffraction Peaks for Form E Relative Two-theta angle (°)d Space (Å) Intensity (%) 3.119693 28.32131 3.07 5.501205 16.06499 4.737.784643 11.35709 100 11.02865 8.02269 17.75 13.51128 6.55363 7.7714.60025 6.06718 60.61 15.63387 5.6683 23.37 16.60508 5.3389 2.1917.49836 5.06831 47.81 18.35094 4.83472 8.95 19.99003 4.44184 7.720.7282 4.2853 18.3 22.17633 4.00531 47.51 22.91616 3.87765 8.0723.53123 3.77767 20.87 24.20048 3.67469 1.84 24.83745 3.58188 19.1726.06663 3.41569 20.46 26.68317 3.33815 4.14 27.26868 3.26779 3.2427.81331 3.20503 16.02 28.38011 3.14229 18.04 29.48873 3.02663 2.0330.00215 2.976 1.96 31.08283 2.87495 2.63 31.60576 2.82856 4.57 32.055682.78988 3.52 32.57093 2.74692 6.43 33.55435 2.66862 1.21 34.003132.63442 1.26 34.50783 2.59704 1.26 35.38724 2.53449 3.74 36.340732.47015 1.03 37.21299 2.41423 4.79 38.06575 2.36208 2.16 39.361662.28725 1.35 39.76171 2.26515 1.83

Form F

Form F was obtained from equilibration of Form A in IPA at 50° C. Form Fhad a crystalline XRPD pattern as shown in FIG. 23. A SEM picture ofForm F is provided as FIG. 24. TGA and DSC thermograms of Form F areshown in FIG. 25 and FIG. 26, respectively. The TGA weight loss of 14.3wt % corresponded to a broad DSC peak with an onset at around 137° C.and can be attributed to loss of solvent in Form F. The DSC peak with amaximum temperature of 153° C. corresponded to the melt/decomposition.The ¹H-NMR spectrum obtained for the Form F sample was consistent withstructure but contained IPA. See FIG. 27. These observations suggestedthat Form F is an IPA solvate of Compound 1.

A list of X-Ray Diffraction Peaks for Form F is provided below in Table17.

TABLE 17 X-Ray Diffraction Peaks for Form F Relative Two-theta angle (°)d Space (Å) Intensity (%) 4.949402 17.85475 3.12 7.016064 12.59939 22.779.436884 9.37204 100 11.121 7.95627 5.93 11.78187 7.51143 96.32 15.452455.73444 24.74 15.77157 5.61912 14.12 16.99932 5.21596 7.49 17.59225.04149 10.1 18.00623 4.92649 80.55 18.29209 4.85014 53.82 18.9724.67783 13.76 19.74617 4.49614 33.34 19.98908 4.44205 33.31 20.25344.38467 6.37 20.87538 4.25542 48.06 22.38658 3.97146 8.46 22.6346 3.92856.09 23.18467 3.83652 15.28 23.72334 3.75061 15.45 24.35122 3.65531 6.7225.08596 3.54989 6.8 25.36348 3.51168 6.38 25.58845 3.48131 7.7326.40269 3.37577 4.91 26.78918 3.32794 9.12 27.25453 3.27216 5.6527.73582 3.21647 13.8 28.63785 3.11718 6.16 29.18351 3.06012 7.4730.04752 2.97407 8.89 30.39711 2.94066 4.57 30.65469 2.91412 4.0131.24242 2.863 2.33 32.07575 2.79049 9.62 34.12926 2.62715 2.68 34.432382.60256 1.58 35.20737 2.54913 1.97 35.8216 2.50682 2.08 36.53788 2.457274.53 38.49454 2.33868 1.47 38.83843 2.31877 2.01 39.24398 2.29573 1.46

Form G

Form G was obtained from equilibration of Form A in MTBE at 50° C. FormG had a crystalline XRPD pattern as shown in FIG. 28. A SEM picture ofForm G is provided as FIG. 29. TGA and DSC thermograms of Form G areshown in FIG. 30 and FIG. 31, respectively. The TGA weight loss of 8.7wt % corresponded to a broad DSC peak around 147° C. and can beattributed to loss of solvent in Form G. The DSC peak with maximum ofabout 161° C. corresponded to the melt/decomposition. The ¹H-NMRspectrum obtained for the Form G sample consistent with structure butcontained MTBE (see FIG. 32). These observations suggested that Form Gis an MTBE solvate of Compound 1.

A list of X-Ray Diffraction Peaks for Form G is provided below in Table18.

TABLE 18 X-Ray Diffraction Peaks for Form G Relative Two-theta angle (°)d Space (Å) Intensity (%) 4.47243 19.75778 2.2 7.954996 11.11426 17.718.975498 9.85274 21.83 9.866793 8.96463 100 9.994242 8.85059 26.5510.16899 8.69889 7.78 11.63523 7.59947 3.23 11.92189 7.42353 11.8413.47007 6.57358 1.61 14.3726 6.16276 7.5 14.62417 6.05731 9.1 15.27955.79896 53.13 15.86499 5.58625 10.48 16.40464 5.40368 16.53 16.938945.23441 18.87 17.48632 5.07177 25.51 17.748 4.99758 7.4 17.98435 4.9324319.01 18.36435 4.83122 33.34 18.69249 4.74714 22.02 18.81186 4.713385.12 19.3983 4.57598 21.65 19.62571 4.52347 26.03 20.30466 4.37372 4.0320.78484 4.27375 2.19 21.17322 4.19622 29.62 21.57257 4.11944 3.6722.034 4.0342 1.86 22.23484 3.99822 3.05 22.52995 3.94651 6.98 22.851363.89172 33.76 23.3702 3.80648 5.76 23.98599 3.71014 10.82 24.451113.6406 9.87 24.6287 3.61176 5.11 24.98931 3.5634 4.79 25.18471 3.53624.77 25.5561 3.48564 7.07 25.88694 3.43899 8.4 25.99959 3.42719 9.9626.43365 3.37189 3.76 26.90873 3.31342 5.69 27.32137 3.26431 1.1827.61015 3.23082 3.29 27.98048 3.1889 4.2 28.16615 3.1683 2.67 28.750343.10524 2.95 29.37178 3.04093 5.27 29.87498 2.99085 1.05 30.223852.95712 0.8 30.78081 2.90487 4.66 31.43885 2.84555 9.53 31.75021 2.818364.58 32.79725 2.73074 1.58 33.21187 2.69759 2.01 34.41132 2.60626 2.3734.90557 2.57048 0.63 35.666 2.5174 2.35 36.10336 2.4879 2.17 38.185292.35691 2.36 38.91115 2.3146 1.7

Form H

Form H was obtained from of Form A in EtOH/water (1:1), EtOH, or EtOAcat 50° C. Form H had a crystalline XRPD pattern as shown in FIG. 33. TGAand DSC thermograms of Form H are shown in FIG. 34 and FIG. 35,respectively. The TGA thermogram weight loss of 6.5 wt % corresponded tobroad DSC peak around 163° C. and can be attributed to loss of solventin Form H. The DSC peak with onset temperature of about 179° C. andmaximum of about 187° C. corresponded to the melt/decomposition. Thetheoretical EtOH content of a mono-solvate of Compound 1 is 7.5%,corresponding to the TGA weight loss observed. These observationssuggested that Form H is a solvate or hydrate of Compound 1. Formtransfer experiment showed that exposing Form H above 20% RH resulted inForm A.

A list of X-Ray Diffraction Peaks for Form H is provided below in Table19.

TABLE 19 X-Ray Diffraction Peaks for Form H Relative Two-theta angle (°)d Space (Å) Intensity (%) 6.13687 14.40231 13.53 7.685174 11.50386 84.298.896353 9.94022 56.97 10.2724 8.61156 100 10.85454 8.15098 4.9411.31227 7.82217 7.69 11.58221 7.64046 7.47 13.71878 6.45497 5.3214.39424 6.15355 8.65 14.97271 5.91708 7.68 15.23389 5.81622 26.0615.36865 5.76552 20.38 15.59046 5.68399 26.21 15.86336 5.58681 4.9616.90296 5.24548 16.49 17.21535 5.15099 21.58 17.74632 4.99391 18.4818.23822 4.86434 76.87 18.6664 4.75372 3.76 19.42537 4.56966 10.4519.62624 4.52335 22.96 20.59336 4.31305 5.08 20.90294 4.24987 11.5521.4434 4.14396 42.91 22.48423 3.95443 8.36 23.20369 3.83342 8.1223.69494 3.75505 2.99 24.58745 3.62072 7.34 24.91904 3.57329 22.5825.58037 3.48239 10.14 25.90445 3.43671 4.73 26.15241 3.4075 4.0526.90531 3.31384 5.27 27.42283 3.25246 7.59 28.13099 3.17218 10.8728.3945 3.14334 19.67 28.96609 3.0826 6.69 29.38076 3.04003 1.7331.09366 2.87635 7.22 32.20206 2.77984 7.9 33.11108 2.70557 3.11 34.09492.62972 1.71 34.72511 2.58342 1.58 35.26616 2.54502 1.97 37.331362.40884 2.19 38.63271 2.33064 3.31

Form I

Form I was obtained from cooling recrystallization of Form A in MeCN.Form I had a crystalline XRPD pattern as shown in FIG. 36. TGA and DSCthermograms of Form I are shown in FIG. 38 and FIG. 39, respectively.The TGA weight loss of 2.3 wt % corresponded to a broad DSC peak around75° C. and can be attributed to loss of solvent in Form I. The DSC peakwith onset temperature of about 173° C. and maximum of about 183° C.corresponded to the melt/decomposition. Form transfer experiment showedthat slurry with MeCN at RT resulted in Form C. These observationssuggested that Form I is a solvate or a hydrate of Compound 1.

A list of X-Ray Diffraction Peaks for Form I is provided below in Table20.

TABLE 20 X-Ray Diffraction Peaks for Form I Relative Two-theta angle (°)d Space (Å) Intensity (%) 5.228867 16.90109 7.86 5.454234 16.20324 15.456.316221 13.99375 100 8.610236 10.26988 13.9 9.260653 9.54999 5.0910.44823 8.46702 7.58 10.92414 8.09921 4.42 11.48793 7.70296 8.2211.94335 7.41024 4.88 12.63676 7.00513 1.81 15.67971 5.65184 24.7616.61849 5.33462 20.31 17.29023 5.12885 11.91 18.14668 4.88867 24.5818.69751 4.74588 11.16 19.02843 4.66408 7.63 20.04536 4.4297 21.3920.88708 4.25306 5.97 21.96666 4.04642 19.84 22.48101 3.95499 4.4723.28344 3.82047 1.5 24.09921 3.69296 7.58 24.61163 3.61722 1.6925.41945 3.50407 4.04 26.40632 3.37531 8.22 27.56053 3.23653 9.4628.37724 3.14521 1.21 29.60591 3.01742 1.4 30.97981 2.88666 1.6931.64884 2.82715 1.68 32.10926 2.78535 2.02 33.24588 2.69491 1.9633.93032 2.64209 0.94 35.26857 2.54485 1.03 35.87462 2.50324 1.2938.49128 2.33887 0.67 35.42 2.5341 0.6 36.56 2.4577 0.5 37.67 2.3880 1.1

Amorphous Solid Free Base

An amorphous solid of Compound 1 was obtained from most evaporationexperiments at room temperature or 50° C., as shown in Table 6.

The amorphous solid had an XRPD spectrum as shown in FIG. 40. DSCthermogram of the amorphous solid sample is shown in FIG. 41. Theamorphous solid has a glass transition temperature of approximately 84°C. The DVS Isotherm plot of the amorphous solid is shown in FIG. 43A. Areversible weight change of about 3.5% was observed between 10 and 50%RH.

TABLE 21 Summary of Form Conversion Experiments Starting Form SolventConditions Resulting Form A none RT and ambient RH (~20- A 30%) for 24hrs A none RT and 0% RH 6 days B + A A none 40° C. and vacuum oven for B48 hrs B none RT and ambient RH (~20- A 30%) for 5 days C none RT andambient RH (~20- A 30%) for 5 days D none RT and ambient RH (~20- D 30%)5 days F none RT and ambient RH (~20- F 30%) 5 days G none RT andambient RH (~20- G 30%) 5 days H none RT and ambient RH (~20- A 30%) 5days I none RT and ambient RH (~20- I 30%) 5 days

Solid Forms Analytical Methods—Citrate Salt Forms

A polymorph screen of the citrate salt Compound 1 was performed toinvestigate whether different solid forms could be generated undervarious conditions, such as different solvents, temperature and humiditychanges.

The solvents used in the polymorph screen were either HPLC or reagentgrade, including acetonitrile (MeCN), MeCN/water (1:1), n-butanol(n-BuOH), absolute ethanol (EtOH), ethanol/water (1:1), methanol (MeOH),2-propanol (IPA), ethyl acetate (EtOAc), methyl acetate (MeOAc),dichloromethane (DCM), methyl ethyl ketone (MEK), methyl t-butyl ether(MTBE), heptane, toluene, methyl acetate (MeOAc), isopropyl acetate(IPAc), methyl isobutyl ketone (MIBK), 2-methyltetrahydrofuran(2-MeTHF), 1,4-dioxane, tetrahydrofuran (THF), THF/water (1:1), water,dimethyl sulfoxide (DMSO), dimethylacetamide (DMA, DMAc), andN-methylpyrrolidone (NMP).

A weighed sample of Compound 1 citrate was treated with a known volumeof a test solvent. The resulting mixture was agitated for 1 day at roomtemperature. If all of the solids appeared to be dissolved by visualinspection, the estimated solubility was calculated based on the totalvolume of solvent used to give a complete solution. If solids werepresent, a known volume of filtrate was evaporated to dryness and theweight of the residue was measured to estimate the solubility.

All of the solid samples generated in the polymorph screen were analyzedby XRPD. XRPD analysis was conducted on a PANalytical Empyrean X-raypowder diffractometer using Cu Kα radiation at 1.54 Å.

The PANalytical Empyrean instrument was equipped with a fine focus X-raytube. The voltage and amperage of the X-ray generator were set at 45 kVand 40 mA, respectively. The divergence slits were set at 1/16° and ⅛°,and the receiving slit was set at 1/16°. Diffracted radiation wasmeasured using a Pixel 2D detector. A theta-two theta continuous scanwas set at step size 0.013 or 0.026 from 3° to 40° 2θ with samplespinning rate at 4. A sintered alumina standard was used to check thepeak positions.

DSC analyses were performed on a TA Discovery Differential Scanningcalorimeter. Indium was used as the calibration standard. Approximately1-5 mg of sample was placed into a DSC pan. The sample was heated undernitrogen at a rate of 10° C./min, up to a final temperature of 260° C.Melting points were reported as the extrapolated onset temperatures.

TGA analyses were performed on a TA Discovery ThermogravimetricAnalyzer. Approximately 2-10 mg of accurately weighed sample was placedon a pan and loaded into the TGA furnace. The sample was heated undernitrogen at a rate of 10° C./min, up to a final temperature of 300° C.

Morphology analysis of the samples was carried out on an Evex Mini-SEM.Small amounts of samples were dispersed on a sample holder, coated withgold using an Evex Mini Au Sputter Coater, and imaged with 500× to 1000×magnification.

Hygroscopicity was determined on a Surface Measurement Systems DVS. Asample size of 5-20 mg was loaded into the DVS instrument sample pan andthe sample was analyzed on a DVS automated sorption analyzer at roomtemperature. The relative humidity was increased from 0% to 90% RH at10% RH step, then decreased in a similar manner to accomplish a fulladsorption/desorption cycle.

¹H NMR spectra were obtained on a Bruker 300 MHz NMR spectrometer.Samples were dissolved in DMSO-D⁶ and analyzed with 128 scans.

Solubility of Form A and Form B in selected organic solvents wasdetermined by mixing the individual solid forms with selected solventsat room temperature. Aliquots were obtained at multiple time points (18hrs, 4 days, 8 days or 12 days), filtered, and quantified by an HPLCmethod. The recovered solids were analyzed by XRPD to confirm the solidforms.

Equilibration/Slurry and Evaporation Experiments

Equilibration and evaporation experiments at room temperature and 50° C.were carried out by adding an excess of Compound 1 citrate solid to upto 1 mL of a test solvent. The resulting mixture was agitated for 1 dayat room temperature and 1 day at 50° C. separately. Upon reachingequilibrium, the saturated supernatant solution was removed, filteredusing 0.45 μm PTFE filters and allowed to evaporate in an open vialunder nitrogen at room temperature and 50° C., respectively. The solidresulting from the equilibration was isolated and air-dried beforeanalysis.

Anti-Solvent Recrystallization and Cooling Recrystallization Experiments

For cooling recrystallization, each of the selected solvents wassaturated with Compound 1 citrate at 65° C. The solvents includedMeCN/water (1:1), EtOH, EtOH/water (1:1), MeOH, THF/water (1:1) and THF.The solution was stirred for 10 minutes, filtered using a 0.45 μm PTFEsyringe filter, and then cooled to −15° C. by placing the vials into afreezer. The solid resulting from the recrystallization was isolated andair-dried before analysis.

For anti-solvent recrystallization, the selected solvent DMA wassaturated with Compound 1 citrate material at the room temperature. Oncethe solid was completely dissolved, a portion of the solution wasfiltered into a vial containing a selected anti-solvent (MeCN, MeOH,heptane, EtOAc, toluene and water). The mixture was cooled to −15° C.and 4° C. by placing the vials into a freezer or a refrigerator. Thesolid resulting from the recrystallization was isolated and air-driedbefore analysis.

Summary of Polymorphic Forms

Two crystalline forms for Compound 1 citrate salt were found during thispolymorph screen study. The stack plot of XRPD patterns for these formsare shown in FIG. 44, and the physical characteristics are summarized inTable 29.

Form Y

Form Y was obtained from dissolving Compound 1 starting material in 5Vol Acetone @ 25 C. About 1.15 eq citric acid in water (˜0.2 M) wascharged into the batch to form the Compound 1 citrate salt. The Compound1 citrate salt was aged at 25° C. until the mother liquor concentrationwas below 1 mg/ml. The slurry was filtered off and washed using ˜4 vol(1:1) Acetone/H₂O to wash the cake. The cake was dried in a vacuum ovenat 50° C. until no acetone was detected by NMR.

Approximate solubility of Compound 1 citrate Form Y in various solventsat ambient temperature was estimated as described above. The results aresummarized in Table 22. Compound 1 citrate Form Y was found to be mostsoluble (>50 mg/mL) in DMSO, DMA and NMP. Compound 1 citrate Form Yshowed some solubility (>20 mg/mL) in THF/water, (>5 mg/mL) in THF, (>3mg/mL) in MeCN/water (1:1) and MeOH, (>2 mg/mL) in 1,4 dioxane. Compound1 citrate Form Y showed low solubility (<1-2 mg/mL) in all othersolvents tested, including Acetone, n-BuOH, MeCN, EtOH, EtOH/water(1:1), IPA, EtOAc, MeOAc, DCM, MTBE, MEK, heptane, toluene, 2-MeTHF andwater.

TABLE 22 Approximate Solubility of Compound 1 citrate Form Y at RoomTemperature Approximate Solubility Solvent (mg/mL) Acetone <1 MeCN <1MeCN/water (1:1) <4 n-BuOH <1 EtOH <2 EtOH/water (1:1) <3 EtOAc <1Heptane <1 IPA <1 DCM <1 MeOAc <1 MeOH <4 MTBE <1 MEK <1 Toluene <1 THF<6 THF/water (1:1) <23 water <1 1,4-dioxane <3 MIBK <1 IPAc <1 2-MeTHF<2 DMA >50 NMP >50 DMSO >50

Equilibration and evaporation experiments were performed at roomtemperature and 50° C. using Compound 1 citrate Form Y as startingmaterial, as described above. The results are summarized in Table 23.Equilibration in MeOH and MeCN/water at 50° C. afforded a unique form,designated as Citrate Salt Form Z. All other equilibration experimentsafforded Compound 1 citrate Form Y or Compound 1 citrate Form Y mixedwith Compound 1 citrate Form Z. Due to relatively low solubility, mostevaporation experiments didn't afford analyzable solid. Evaporation fromEtOH and EtOH/water afforded mixture of Compound 1 citrate Forms Y andZ. Solids from MeOH evaporation afforded Compound 1 citrate Form Z.

TABLE 23 Summary of Equilibration and Evaporation Results. Form by XRPDSolvent EQ at RT EV at RT EQ at 50° C. EV at 50° C. Acetone Y — Y — MeCNY — Y — MeCN/water Z + Y — Z — n-BuOH Y — Y — EtOH Y — Z + Y Z + YEtOH/water Z + Y Y + Z Z + Y Y + Z EtOAc Y — Y — Heptane Y — Y — IPA Y —Y — DCM Y — Y — MeOAc Y — Y — MeOH Z Z Z Z MTBE Y — Y — MEK Y — Y —Toluene Y — Y — THF Y Y Y Y THF/water Y Y Y Z + Y water Y — Y —1,4-dioxane Y — Y — MIBK Y — Y — IPAc Y — Y — 2-MeTHF Y — Y — n/a: noexperiment —: not analyzable. *: significant degradation occurred.

Cooling recrystallization experiments were performed as described above.The solvents included MeCN/water (1:1), EtOH/water (1:1), THF/water(1:1), EtOH, MeOH and THF. The results are summarized in Table 24. Thesolids obtained from THF and THF/water were confirmed to be Compound 1citrate Form Y. The solids obtained from MeOH and MeCN/water wereconfirmed to be Compound 1 citrate Form Z. The solids obtained from EtOHand EtOH/water were confirmed to be mixture of Compound 1 citrate FormsY and Z.

TABLE 24 Results from Cooling Recrystallization Solvent Cooling ProfileForm by XRPD MeCN/water (1:1) 65 to −15° C. Z EtOH 65 to −15° C. Z + YEtOH/water (1:1) 65 to −15° C. Z + Y MeOH 65 to −15° C. Z THF 65 to −15°C. Y THF/water (1:1) 65 to −15° C. Y

Recrystallizations with anti-solvents were performed as described above.DMA was used as the primary solvent. MeCN, MeOH, heptane, EtOAc, tolueneand water were used as anti-solvents. The results are summarized inTable 25. Using XRPD, the solids obtained from DMA/MeCN, DMA/MeOH andDMA/water were confirmed to be Compound 1 citrate Form Z. The solidsobtained from DMA/EtOAc were confirmed to be Compound 1 citrate Form Yand the solids obtained from DMA/toluene were confirmed to be a mixtureof Compound 1 citrate Forms Y and Z. Precipitation was not observed fromthe DMA/heptane recrystallization experiment.

TABLE 25 Results from Anti-Solvent Recrystallization Primary Solventsolvent Anti-Solvent Ratio Cooling profile Form by XRPD DMA MeCN 1:15 RTto −15° C. Z DMA MeOH 1:15 RT to −15° C. Z DMA heptane 1:15 RT to −15°C. — DMA EtOAc 1:15 RT to −15° C. Y DMA toluene 1:15 RT to −15° C. Y + ZDMA water 1:15 RT to 4° C. Z RT: room temperature —: no precipitation

Form Y was designated as the crystalline form of the DSD sample used asthe starting material for this screen. Form Y has a crystalline XRPDpattern as shown in FIG. 45. The SEM picture is shown in FIG. 46. TGAand DSC thermograms of Form Y are shown in FIG. 47 and FIG. 48,respectively. No TGA weight loss was observed up to 150° C. for Form Y.Small additional weight loss was observed up to the melting temperatureof Form Y. The DSC thermogram showed a melting event with an onsettemperature of 213° C. and a maximum of 217° C. Form Y is a slightlyhygroscopic, with about 2.1% w/w water uptake between 0 and 90% RH. The¹H NMR spectrum is consistent with the structure of a citrate salt, withabout 0.2% w/w of residual acetone (FIG. 50). The citric acid contentwas 25.1 wt % as determined by HPLC, consistent with a 1:1 salt (withtheoretically 25.2 wt % of citric acid). These observations suggest FormY is most likely an anhydrate of Compound 1 citrate.

The stability of Form Y was further characterized by compression testand form transfer experiments. Upon application of 2000-psi pressure forabout 1 minute, the material was still Form Y (FIG. 51A and FIG. 51B).

TABLE 26 HPLC Solubility of Compound 1 citrate Form Y and Form Z inSelected Solvents at Room Temperature. Solubility (mg/mL) Form Solvent18 hours 4 days 8 days 12 days Y Acetone 0.91 — — — Z Acetone 1.30 — — —Y EtOH 2.27 — — — Z EtOH 1.35 — — — Y MeOAc 0.15 0.20 — 0.20 Z MeOAc0.28 0.27 — 0.32 Y 2-MeTHF 1.48 — 1.25 — Z 2-MeTHF 1.89 — 1.91 — —: nottested

Note: all solids recovered from the solubility tests remained as thestarting form by XRPD.

A list of X-Ray Diffraction Peaks for Form Y is provided below in Table27.

TABLE 27 X-Ray Diffraction Peaks for Form Y Relative Two-theta angle (°)d Space (Å) Intensity (%) 4.783092 18.47519 77.84 6.5819 13.42948 22.249.59256 9.22029 77.51 13.60691 6.50778 15.93 14.38278 6.15842 8.3915.38972 5.75768 42.05 15.96684 5.55084 21.08 16.88841 5.24996 35.5818.02213 4.92218 8.66 18.85463 4.70668 90.57 19.24503 4.61208 10019.93024 4.45503 24.22 20.10453 4.4168 17.8 20.90504 4.24944 31.3721.84462 4.06875 9.14 22.41502 3.96648 11.55 22.68635 3.91965 13.8123.18753 3.83605 4.81 23.41127 3.79675 3.74 23.96094 3.71088 13.5424.11531 3.68748 15.13 24.34514 3.65318 6.5 25.08466 3.55007 5.5226.69176 3.33986 14.16 27.00945 3.3013 11.93 27.91142 3.19663 4.4928.54004 3.12764 26.41 29.0111 3.07791 8.77 29.56958 3.02104 2.9630.15901 2.96333 3.37 30.44673 2.93355 6.11 30.78504 2.90208 4.7131.14674 2.87157 5.23 31.59191 2.83212 5.94 32.30739 2.77101 7.9633.13792 2.7012 2.5 33.53919 2.67201 5.44 33.98455 2.638 8.13 34.594262.59075 2.82 35.05464 2.55989 5.67

Form Z

Compound 1 citrate Form Z was generated by equilibration experiment inMeOH and MeCN/water (1:1) at 50° C. and various recrystallizationexperiments, including cooling recrystallization from MeCN/water, andanti-solvent recrystallization from DMA/MeCN, DMA/MeOH and DMA/water.Form Z has a crystalline XRPD pattern as shown in FIG. 52. The SEMpicture is shown in FIG. 53. TGA and DSC thermograms of Form Z are shownin FIG. 54 and FIG. 55, respectively. No significant TGA weight loss wasobserved for Form Z up to 150° C. Additional weight loss (up to a fewpercent) was observed up to the melting temperature of Form Z. The DSCthermogram showed a melting event with an onset temperature of 217° C.and a maximum of 221° C. Form Z is slightly hygroscopic, with about 1.6%w/w water uptake between 0 and 90% RH. The ¹H NMR spectrum for the FormZ sample out equilibration in MeCN/water at 50° C. is consistent withthe structure of Compound 1 citrate salt, with about 1.0% w/w ofresidual MeCN (FIG. 57). The citric acid content was 24.6 wt % asdetermined by HPLC, consistent with a 1:1 salt (with theoretically 25.2wt % of citric acid). These observations suggest Form Z is most likelyan anhydrate of Compound 1 citrate.

TABLE 28 Experiments to Generate Materials for Characterization SolventExperimental Conditions Form by XRPD MeOH Slurry starting with Form Y, Zshaking at 50° C. for 1 day MeCN/Water(1:1) Slurry starting with Form Y,Z shaking at 50° C. for 1 day

Further drying study was performed to understand the weight lossobserved above 150° C. Aliquots of Form Z sample was dried in KF oven(with N₂ sweep) at 150 and 180° C., respectively. Citric acid content ofthe recovered solids was 24.2 and 17.8 wt %, respectively, suggestingloss of citric acid. Residual solvent in the dried samples were alsosignificantly lower than the “as-is” Form Z sample.

The stability of Form Z was further characterized by compression testand form transfer experiments. Upon application of 2000-psi pressure forabout 1 minute, the material was still Form Z (FIGS. 62A-B).

TABLE 29 Summary of Characterization Data for Compound 1 Citrate SaltPolymorphs Representative DSC peak DVS or other Form Descriptionconditions (° C.) TGA loss (wt %) comments Y anhydrate starting materiallot 213 0.0 (up to 150° C.) ~2.1 wt % water 8153-002; (onset) uptake upto 90% recrystallization in RH; THF and THF/water; anti-solventrecrystallization in DMA/EtOAc Z anhydrate recrystallization or 217 0.1(up to 150° C.) ~1.6 wt % water equilibration in MeOH (onset) uptake upto 90% and MeCN/water; RH; anti-solvent recrystallization in DMA/MeCN,DMA/MeOH, DMA/water

A list of X-Ray Diffraction Peaks for Form Z is provided below in Table30.

TABLE 30 X-Ray Diffraction Peaks for Form Z. Relative Two-theta angle(°) d Space (Å) Intensity (%) 4.648808 19.00855 146.09 6.594748 13.40334240.86 9.398551 9.41017 2485.09 13.10081 6.75803 615.48 14.08974 6.28584474.71 15.33978 5.77631 478.60 15.62982 5.66976 353.57 17.40622 5.09493817.10 18.80665 4.71858 6793.99 18.95349 4.68235 3368.79 19.8844 4.4652486.73 20.41888 4.34951 138.16 21.09284 4.21203 552.13 21.89109 4.06021539.72 22.16856 4.01002 861.32 22.70611 3.91629 337.89 23.51471 3.78342774.70 23.9067 3.72226 103.10 25.16965 3.53828 208.07 26.30357 3.38827243.12 26.80383 3.32615 737.22 27.7519 3.21464 133.59 28.29834 3.1538359.75 28.72703 3.1077 950.99 29.79863 2.99834 103.84 31.19139 2.86756240.17 31.88523 2.80673 82.73 32.60755 2.74619 220.23 33.73557 2.6569113.82 35.08647 2.55764 120.37 35.93019 2.49949 177.27 37.39344 2.4049835.78 37.99067 2.36853 47.79

The thermodynamic relationship between the two forms was exploredthrough form conversion experiments (Table 31). Competitive slurriesstarting with mixtures of Forms Y and Z resulted in solvent specificresults. Form Y resulted from slurries in THF at room temperature andfrom slurries in THF and EtOAc at 50° C. Form Z resulted from slurriesin EtOH, water, MEK, and MeCN at both room temperature and 50° C.

The thermodynamic relationship between the two forms was furtherexplored through solubility experiments (Table 26). These experimentswere designed to determine whether results from competitive slurrieswere due to the different dissolution/growth kinetics of each form in aspecific solvent or the overall thermodynamics. As shown in Table 26,the solubility of Form Z was lower than that of Form Y in EtOH, whilethe solubility of Form Y was lower in acetone, MeOAc, and 2-MeTHF. Theseresults appear consistent with observations from competitive slurries,suggesting that the dissolution/growth kinetics was not the cause forthe solvent specific form conversion.

TABLE 31 Summary of Form Transfer Experiments Starting Temperature/Resulting Form(s) Solvent Condition Form(s) Y + Z Slurry in MeCN RT, 5days Y Y + Z Slurry in EtOH RT, 5 days Z Y + Z Slurry in EtOAc RT, 5days Y + Z Y + Z Slurry in Toluene RT, 5 days Z + Y Y + Z Slurry in THFRT, 5 days Y Y + Z Slurry in Water RT, 5 days Z Y + Z Slurry in MEK RT,5 days Z Y + Z Slurry in MeCN RT, 10 days Z Y + Z Slurry in EtOH RT, 10days Z Y + Z Slurry in EtOAc RT, 10 days Y + Z Y + Z Slurry in TolueneRT, 10 days Z + Y Y + Z Slurry in THE RT, 10 days Y Y + Z Slurry in MEKRT, 10 days Z Y + Z Slurry in MeCN 50° C., 4 days Z Y + Z Slurry in EtOH50° C., 4 days Z Y + Z Slurry in EtOAc 50° C., 4 days Y Y + Z Slurry inToluene 50° C., 4 days Z + Y Y + Z Slurry in THE 50° C., 4 days Y Y + ZSlurry in MEK 50° C., 4 days Z

Solid Forms Analytical Methods—HCl Salt Forms

A polymorph screen of Compound 1 was performed to investigate whetherdifferent solid forms could be generated under various conditions, suchas different solvents, temperature and humidity changes.

The starting material was generated by dissolving Compound 1 in 10 volMeOH at 25-30° C. Then 1.10 equiv HCk in MeOH (˜1.25 M) was charged intothe batch to form Compound 1 HCl Salt. Constant vacuum distillation tosolvent switch from MeOH to EtOAc (˜30-35 vol) was performed and thebatch temperature was maintained at 25-35° C. The slurry was filteredoff, and ˜5 vol (1:1) EtOAc was used to wash the cake, which was driedin a vacuum oven at 50° C.

Starting material is in relatively low crystallinity with a weight lossof 1.1% wt % up to 100° C. in TGA and one melting peak at 238.5° C.(onset temperature) in DSC.

A mass change of 3.6 wt % was observed for starting material from 0% RHto 95% RH at 25° C. The sample is moderately hygroscopic

The theoretical Cl content for a 1:1 HCl salt is 5.84 wt %. The Clcontent of the HCl salt of Compound 1 was 5.70 wt %.

Solubility of the starting mater in selected organic solvents wasdetermined by mixing with selected solvents at room temperature.

TABLE 32 Approximate solubility of anhydrate/amorphous starting materialApproximate Approximate Solvent Solubility (mg/mL) Solvent Solubility(mg/mL) MeOH S > 31 DCM S < 1.6 EtOH S > 28 CHCl3 1.3 < S < 2.0 IPA5.5 > S > 7.3 Toluene S < 1.4 Acetone 2.4 < S < 2.7 Heptane S < 1.7 MIBKS < 1.5 DMAc S > 18 EtOAc S < 1.5 DMSO S > 22 IPAc S < 1.3 NMP S > 24THE 2.3 < S < 3.4 H2O S < 1.0 2-MeTHF S < 0.9 n-BuOH 4.8 < S < 6.31,4-dioxane 1.5 < S < 1.9 MeOAc S < 0.9 MTBE S < 1.0 MEK 1.8 < S < 2.3MeCN 1.1 < S < 1.5

Eight crystalline forms were found during the polymorph screen, andtermed HCl Salt Form 1 through HCl Salt Form 8 herein. Generalcharacteristics of the crystalline forms are provided in Table 33.

TABLE 33 Polymorph Characterization DSC endo TGA Representative peaks (%wt loss Form Description Conditions (onset ° C.) up to ° C.) DVSAnhydrate Starting material 238 1.1 (100° C.) ~3.6% wt (mixed with gain@ amorphous) 95% RH; 1 solvate (IPA) EV@ RT in IPA 102, 146, . . . 6.6(140° C.) 16.8 (205° C.) 2 unknown LVD in IPA/toluene 151, . . . 2.6(140° C.) 16.8 (200° C.) 3 unknown LVD in n-BuOH/ 153, . . . 2.3 (140°C.) heptane 13.2 (210° C.) 4 solvate LVD in MeOH/IPAc 94, 161{circumflexover ( )}, 219 4.5 (140° C.) 5 solvate or SVD in DMF 104 . . . 162, 1924.2 (140° C.) hydrate 6 Pentahydrate Slurry anhydrate in 53, 201 11.6(100° C.) 0.1N HCl at RT 7 monohydrate 1 Slurry anhydrate in 80, 215 3.8(100° C.) ~4.5% wt water at RT gain @ 95% RH, w/ hysterisis EV =evaporation RH = Relative Humidity RT = Room Temperature LVD = LiquidVapor Diffusion SVD = Solid Vapor Diffusion

HCl Salt Form 1

HCl Salt Form 1 has a crystalline XRPD pattern as shown in FIG. 66. TGAand DSC thermograms of HCl Salt Form 1 are shown in FIG. 67. The DSCthermogram showed multiple thermal events one event having an onsettemperature of about 102° C. and maximum of about 114° C., attributed todehydration and a second having an onset temperature of about 146° C.and maximum of about 181° C., corresponding to melt/decomposition. TGAweight loss was 6.6% weight loss before 140° C. and another 10% weightloss before 205° C. HCl Salt Form 1 was obtained from slow evaporationwith IPA. HCl Salt Form 1 is an IPA solvate of the HCl salt of Compound1.

A list of X-Ray Diffraction Peaks for HCl Salt Form 1 is provided belowin Table 34.

TABLE 34 X-Ray Diffraction Peaks for HCl Salt Form 1. Relative Two-thetaangle (°) d Space (Å) Intensity (%) 7.043121 12.55104 17.58 8.75065310.10539 15.35 9.852726 8.9774 100 11.47008 7.7149 17.12 12.158037.27987 11.47 12.43821 7.1165 66.17 14.72797 6.01485 19.95 16.347895.42231 25.48 16.7059 5.30251 3.34 17.47299 5.07561 64.11 17.912314.95211 76.6 18.2135 4.87089 8.18 18.60715 4.76872 4.99 19.16845 4.6303342.87 19.35014 4.58726 15.37 19.71049 4.5042 86.74 19.89059 4.4638256.77 20.26675 4.38181 3.67 20.99857 4.23073 21.73 21.24423 4.18236 6.5921.89672 4.05918 9.98 22.69272 3.91857 11.28 22.87039 3.88853 12.8123.71329 3.75218 11.12 24.62266 3.61563 16.07 24.95768 3.56785 21.9725.58616 3.48162 20.79 26.26022 3.39376 11.72 26.54722 3.35772 6.8926.79783 3.32688 12.32 27.24697 3.27306 20.4 27.60085 3.22921 3.0528.22986 3.16129 10.88 28.68693 3.11195 16.16 29.10407 3.06576 4.8129.63481 3.01454 12.04 30.27581 2.95216 2.91 30.83935 2.89949 6 31.231232.864 4.49 31.67746 2.82466 2.46 32.26273 2.77475 23.06 32.84882 2.726570.93 33.26344 2.69352 3 33.97406 2.63661 2.64 34.30005 2.61446 435.31073 2.54191 1.4 36.2495 2.4782 2.54 36.95941 2.43222 1.43

HCl Salt Form 2

HCl Salt Form 2 has a crystalline XRPD pattern as shown in FIG. 68. TGAand DSC thermograms of HCl Salt Form 2 are shown in FIG. 69. The DSCthermogram showed a broad thermal event with an onset of about 151° C.,attributed to melting, decomposition, and disproportionation. TGA weightloss was 2.6% weight loss before 140° C. and another 14% weight lossbefore 200° C. HCl Salt Form 2 was obtained from liquid vapor diffusionwith IPA/Toluene.

A list of X-Ray Diffraction Peaks for HCl Salt Form 2 is provided belowin Table 35.

TABLE 35 X-Ray Diffraction Peaks for HCl Salt Form 2. Relative Two-thetaangle (°) d Space (Å) Intensity (%) 5.513929 16.02795 39.15 5.81835815.19 8.38 8.983458 9.84403 56.91 9.766082 9.05685 50.75 9.9024498.93243 56.24 10.72871 8.24629 18.43 10.99295 8.04866 10.95 12.260587.21921 27.19 12.57368 7.04013 31.94 13.97997 6.33495 7.85 16.738095.29677 95.21 18.12082 4.89559 22.15 19.03805 4.66175 100 19.522234.54721 7.52 19.94127 4.44891 7.08 20.14258 4.40854 10.91 20.766434.27749 13.97 21.45635 4.14149 23.47 21.7551 4.08529 6.27 22.041314.03288 18.86 22.8351 3.89446 39.99 23.29623 3.8184 9.22 23.986633.71004 5.4 24.28375 3.66531 11.47 24.61877 3.61619 7.17 24.93034 3.571713.85 25.31155 3.51876 6.55 25.98875 3.42859 4.92 26.50429 3.36028 3.7226.80338 3.32621 16.19 27.02674 3.29922 8.76 27.60649 3.22857 2.2528.3911 3.1437 2.89 29.16521 3.062 11.63 29.73827 3.00429 15.98 30.737512.90886 3.83 32.9862 2.71552 7.23 35.1076 2.55615 3.37

HCl Salt Form 3

HCl Salt Form 3 has a crystalline XRPD pattern as shown in FIG. 70. TGAand DSC thermograms of HCl Salt Form 3 are shown in FIG. 71. The DSCthermogram showed a broad thermal event at 153° C. (onset), attributedlikely to melting, decomposition, and disproportionation. TGA weightloss was 2.3% weight loss before 140° C. and another 11% weight lossbefore 210° C. HCl Salt Form 3 was obtained from multiple conditionsrelated to n-BuOH.

A list of X-Ray Diffraction Peaks for HCl Salt Form 3 is provided belowin Table 36.

TABLE 36 X-Ray Diffraction Peaks for HCl Salt Form 3. Relative Two-thetaangle (°) d Space (Å) Intensity (%) 5.479806 16.12768 100 6.48787713.62388 23.65 7.727824 11.44046 2.98 10.02748 8.82133 5.09 10.456058.45371 4.84 10.87943 8.13239 20.4 12.93306 6.8453 18.49 14.841695.96902 7.58 15.91137 5.56546 10.26 16.23176 5.46084 80.27 18.300414.84795 5.46 18.92492 4.68936 9.72 19.44416 4.56529 52.36 20.411974.35097 28.32 20.95774 4.23888 5.68 21.77105 4.08233 18.78 22.209124.00279 22.42 22.48069 3.95504 23.87 24.06092 3.69875 5.4 25.990893.42831 8.79 28.79054 3.10099 1.8 29.8991 2.98849 3.51 32.70738 2.738032.18 39.41492 2.28617 4.53

HCl Salt Form 4

HCl Salt Form 4 has a crystalline XRPD pattern as shown in FIG. 72. TGAand DSC thermograms of HCl Salt Form 4 are shown in FIG. 73. The DSCthermogram showed a desolvation endotherm at 94° C. and a followingexotherm and another endotherm at 219° C. TGA weight loss 4.5% weightloss before 140° C. HCl Salt Form 4 was obtained from liquid vapordiffusion with MeOH/IPAc. HCl Salt Form 4 may be a solvate of the HClsalt of Compound 1.

A list of X-Ray Diffraction Peaks for HCl Salt Form 4 is provided belowin Table 37.

TABLE 37 X-Ray Diffraction Peaks for HCl Salt Form 4. Relative Two-thetaangle (°) d Space (Å) Intensity (%) 7.938752 11.13696 84.85 8.05813810.97223 51.11 8.219399 10.74841 15.24 8.403137 10.52251 48.95 10.824058.17387 4.19 14.02049 6.31673 2.92 15.33837 5.77683 10.12 15.886315.5788 61.42 16.43516 5.38925 2.52 16.80188 5.2768 25.48 17.763914.99314 2.94 18.28634 4.85165 12.92 18.89575 4.69653 35.91 19.144114.63616 100 19.66527 4.51446 24.64 20.25129 4.38512 2.59 21.01279 4.227914.75 21.59817 4.11121 3.83 22.12687 4.01748 1.47 23.30987 3.8162 7.5723.64596 3.76271 13.56 24.92983 3.57177 5.47 25.29449 3.5211 14.9326.25757 3.3941 6.32 27.63529 3.22794 2.52 28.46236 3.136 4.08 29.071773.07163 2.25 29.89888 2.98852 5.3 30.55282 2.92603 5.48 30.85491 2.898064.83 31.85292 2.8095 2.13 32.84365 2.72698 4.22 34.62294 2.59081 1.6536.19538 2.48179 1.87

HCl Salt Form 5

HCl Salt Form 5 has a crystalline XRPD pattern as shown in FIG. 74. TGAand DSC thermograms of HCl Salt Form 5 are shown in FIG. 75. The DSCthermogram showed a desolvation endotherm at about 104° C. and about162° C. followed by a possible melting endotherm with an onset at 192°C. and maximum at 209° C. TGA weight loss was 4.2% weight loss before140° C. HCl Salt Form 5 was obtained from solid vapor diffusion withDMF. HCl Salt Form 5 may be a solvate of the HCl salt of Compound 1. HClSalt Form 5 may be a hydrate of the HCl salt of Compound 1.

A list of X-Ray Diffraction Peaks for HCl Salt Form 5 is provided belowin Table 38.

TABLE 38 X-Ray Diffraction Peaks for HCl Salt Form 5. Relative Two-thetaangle (°) d Space (Å) Intensity (%) 7.937265 11.13905 82.3 8.68907810.17687 44.75 9.993996 8.85081 31.28 11.70052 7.56347 22.93 13.316246.64917 17.53 13.5735 6.51833 6.97 15.06167 5.88233 35.16 15.699895.64462 44.81 17.1975 5.15629 40.42 17.88152 4.95646 9.74 19.864254.46968 100 20.55088 4.32187 46.64 21.33943 4.16391 21.55 23.286083.82004 15.52 24.21503 3.67556 21.33 25.5195 3.49056 11.25 26.962773.30691 55.77 28.52699 3.12904 17.37 29.26391 3.0519 6.89 30.129722.96614 6.25 31.71499 2.81907 13.29 32.19053 2.7808 11.4 34.091242.62999 4.68 35.35318 2.53895 2.83 36.98646 2.4305 2.08 38.75596 2.323512.17

HCl Salt Form 6

HCl Salt Form 6 has a crystalline XRPD pattern as shown in FIG. 76. TGAand DSC thermograms of HCl Salt Form 6 are shown in FIG. 77. The DSCthermogram showed a dehydration endotherm at 88° C. and anotherendotherm with an onset at 201° C. and maximum at 228° C. TGA weightloss was 11.6% weight loss before 100° C. A second run DSC thermogramshowed an endotherm at 89° C. and another endotherm with an onset at211° C. and maximum at 225° C. (FIG. 79). The TGA weight losscorresponded to 15.9% weight loss before 120° C. (FIG. 78). The sampleappeared to contain both surface and crystal water. The theoreticalwater content for a pentahydrate and hexahydrate is 12.9% and 15.1%,respectively. HCl Salt Form 6 was obtained from slurry in 0.1N HCl inwater. HCl Salt Form 6 is a pentahydrate or a hexahydrate of the HClsalt of Compound 1.

A list of X-Ray Diffraction Peaks for HCl Salt Form 6 is provided belowin Table 39.

TABLE 39 X-Ray Diffraction Peaks for HCl Salt Form 6. Two-theta angle(°) d Space (Å) Relative Intensity (%) 7.043121 12.55104 17.58 8.75065310.10539 15.35 9.852726 8.9774 100 11.47008 7.7149 17.12 12.158037.27987 11.47 12.43821 7.1165 66.17 14.72797 6.01485 19.95 16.347895.42231 25.48 16.7059 5.30251 3.34 17.47299 5.07561 64.11 17.912314.95211 76.6 18.2135 4.87089 8.18 18.60715 4.76872 4.99 19.16845 4.6303342.87 19.35014 4.58726 15.37 19.71049 4.5042 86.74 19.89059 4.4638256.77 20.26675 4.38181 3.67 20.99857 4.23073 21.73 21.24423 4.18236 6.5921.89672 4.05918 9.98 22.69272 3.91857 11.28 22.87039 3.88853 12.8123.71329 3.75218 11.12 24.62266 3.61563 16.07 24.95768 3.56785 21.9725.58616 3.48162 20.79 26.26022 3.39376 11.72 26.54722 3.35772 6.8926.79783 3.32688 12.32 27.24697 3.27306 20.4 27.60085 3.22921 3.0528.22986 3.16129 10.88 28.68693 3.11195 16.16 29.10407 3.06576 4.8129.63481 3.01454 12.04 30.27581 2.95216 2.91 30.83935 2.89949 6 31.231232.864 4.49 31.67746 2.82466 2.46 32.26273 2.77475 23.06 32.84882 2.726570.93 33.26344 2.69352 3 33.97406 2.63661 2.64 34.30005 2.61446 435.31073 2.54191 1.4 36.2495 2.4782 2.54 36.95941 2.43222 1.43

HCl Salt Form 7

HCl Salt Form 7 has a crystalline XRPD pattern as shown in FIG. 80. TGAand DSC thermograms of HCl Salt Form 7 are shown in FIG. 81. The DSCthermogram showed a dehydration endotherm with an onset at 80° C. andmaximum at 110° C. and another endotherm with an onset at 215° C. andmaximum at 233° C. TGA weight loss was 3.8% weight loss before 100° C. Asecond run DSC thermogram showed an endotherm with an onset at 71° C.and maximum at 98° C. and another endotherm with an onset at 209° C. andmaximum at 230° C. (FIG. 83). The TGA weight loss corresponded to 3.4%weight loss before about 90° C. (FIG. 82). The theoretical water contentfor a monohydrate is 2.9%. HCl Salt Form 7 was obtained from waterslurry at RT and air dried for 2 weeks. HCl Salt Form 7 is a monohydrateof the HCl salt of Compound 1.

A list of X-Ray Diffraction Peaks for HCl Salt Form 7 is provided belowin Table 40.

TABLE 40 X-Ray Diffraction Peaks for HCl Salt Form 7. Two-theta angle(°) d Space (Å) Relative Intensity (%) 7.850335 11.2622 40.81 8.09212910.92622 63.33 8.328633 10.61647 44.79 10.76644 8.21748 20.86 13.810366.41237 13.87 14.50773 6.10566 3.83 15.29982 5.7913 36.58 15.600445.67568 5.43 16.15992 5.48495 43.64 16.64025 5.32769 18.84 16.976775.22283 11.15 17.63451 5.02949 11.9 18.33576 4.83869 29.55 18.649244.75805 17.26 19.11633 4.64284 100 19.57 4.53246 32.57 19.73225 4.4992851.13 20.1927 4.39408 5.64 20.74966 4.28091 14.35 21.5192 4.12953 21.422.03008 4.03491 12.02 22.87123 3.88838 10.43 24.00267 3.7076 25.5824.25357 3.66981 24.51 25.1557 3.54021 21.68 26.22969 3.39764 14.8528.35525 3.1476 5.24 29.02731 3.07369 7.17 29.51012 3.02699 9.9430.17205 2.95963 5.92 30.75171 2.90515 13.78 31.15329 2.87098 13.832.53576 2.75208 5.9 33.12518 2.70221 7.24 34.66455 2.5878 6.58 36.30542.47452 5.15

HCl Salt Form 8

HCl Salt Form 8 has a crystalline XRPD pattern as shown in FIG. 85. TGAand DSC thermograms of HCl Salt Form 8 are shown in FIG. 86. The DSCthermogram showed a dehydration endotherm with an onset at 117° C. andmaximum at 146° C. and another endotherm with an onset at 208° C. andmaximum at 221° C. TGA weight loss was 3.2% weight loss before 100° C. Asecond run DSC thermogram showed an endotherm at 148° C. (maximum) andanother endotherm with an onset at 204° C. and maximum at 224° C. (FIG.88). The TGA weight loss corresponded to 3.0% weight loss before 120° C.(FIG. 87). The theoretical water content for a monohydrate is 2.9%. HClSalt Form 8 was obtained from water slurry at 50° C. and air dried for 2weeks. HCl Salt Form 8 is a monohydrate of the HCl salt of Compound 1.

A list of X-Ray Diffraction Peaks for HCl Salt Form 8 is provided belowin Table 41.

TABLE 41 X-Ray Diffraction Peaks for HCl Salt Form 8. Two-theta angle(°) d Space (Å) Relative Intensity (%) 8.103976 10.91027 8.9 9.757069.0652 76.5 10.11246 8.7474 14.63 10.81305 8.17539 6.87 11.07871 7.9865511.12 11.61823 7.61686 3.64 15.67207 5.65457 10.72 16.16669 5.48267 7.7116.85717 5.25962 6.54 17.39887 5.09707 100 17.96866 4.9367 48.7118.67773 4.75086 86.44 19.16066 4.63219 8.88 19.60107 4.5291 14.5421.35427 4.16105 18 22.05148 4.03104 3.73 22.94882 3.87541 14.4 23.843583.73197 13.2 24.24392 3.67125 13.61 25.06726 3.5525 4.2 25.45586 3.499145.79 26.24939 3.39514 71.25 26.70115 3.33871 10.55 28.15429 3.16961 11.328.43738 3.13869 12.07 29.57393 3.02061 3.56 30.49194 2.93173 10.0831.73701 2.8195 10.32 31.99273 2.79754 8.69 33.69307 2.66015 4.2335.57793 2.52343 4.42 36.36242 2.47077 3.1 37.37213 2.4063 4.35

Starting Material HCl Salt Form

The starting material HCl Salt Form has a crystalline XRPD pattern asshown in FIG. 63. TGA and DSC thermograms of starting material HCl SaltForm are shown in FIG. 64. The DSC thermogram showed one endotherm withan onset at about 238° C. and maximum at about 248° C. The TGA weightloss corresponded to about 1.0% weight loss before 100° C. A mass changeof 3.6 wt % was observed for starting material from 0% RH to 95% RH at25° C. The Cl content was 5.70 wt % and is in agreement with thetheoretical Cl content for a 1:1 HCl salt of 5.84 wt %. The sample ismoderately hygroscopic. The starting material HCl Salt Form may be ananhydrate form of the HCl salt of Compound 1.

A list of X-Ray Diffraction Peaks for starting material HCl Salt Form isprovided below in Table 42.

TABLE 42 X-Ray Diffraction Peaks for starting material HCl Salt Form.Two-theta angle (°) d Space (Å) Relative Intensity (%) 5.828535 15.163515.88 7.062807 12.5161 15.18 8.277888 10.68144 26.8 10.14078 8.7230346.8 11.34123 7.80226 54.67 11.60621 7.62472 56.11 12.71036 6.9647334.71 15.51306 5.71217 40.65 16.13662 5.49282 53.48 17.82084 4.9773256.68 19.19574 4.62381 75.92 19.67368 4.51255 50.35 20.53309 4.3255762.22 21.13495 4.20374 100 22.97894 3.8704 30.97 23.97222 3.71224 27.3825.54515 3.48711 19.36 26.33352 3.38448 22.8 27.20446 3.27807 29.0928.42589 3.13994 21.17 31.04672 2.88059 6.68

Cell Assays

Multiplexed Cytotoxicity Assay.

Cells are grown in RPMI1640, 10% FBS, 2 mM L-alanyl-L-Glutamine, 1 mM Napyruvate or a special medium in a humidified atmosphere of 5% CO₂ at 37°C. Cells are seeded into 384-well plates and incubated in a humidifiedatmosphere of 5% CO₂ at 37° C. Compounds are added 24 h post cellseeding. At the same time, a time zero untreated cell plate isgenerated. After a 72 hour incubation period, cells are fixed andstained with fluorescently labeled antibodies and nuclear dye to allowvisualization of nuclei, apoptotic cells and mitotic cells. Apoptoticcells are detected using an anti-active caspase-3 antibody. Mitoticcells are detected using an anti phospho-histone-3 antibody. Compoundsare serially diluted 3.16-fold and assayed over 10 concentrations in afinal assay concentration of 0.1% DMSO from the highest testconcentration of 10 μM. Automated fluorescence microscopy was carriedout using a Molecular Devices ImageXpress Micro XL high-content imager,and images are collected with a 4× objective.

Data Analysis.

Sixteen-bit TIFF images are acquired and analyzed with MetaXpress5.1.0.41 software. Cell proliferation is measured by the signalintensity of the incorporated nuclear dye. The cell proliferation assayoutput is referred to as the relative cell count. To determine the cellproliferation end point, the cell proliferation data output istransformed to percentage of control (POC) using the following formula:

POC=relative cell count(compound wells)/relative cell count(vehiclewells)×100

Relative cell count IC₅₀ is the test compound concentration at 50% ofmaximal possible response relative to the DMSO control. GI₅₀ is theconcentration needed to reduce the observed growth by half. This is theconcentration that inhibits the growth to the level midway betweengrowth in untreated cells and the number of cells seeded in the well(Time zero value). The IC₅₀ values are calculated using nonlinearregression to fit data to a sigmoidal 4 point, 4 parameter One-Site doseresponse model, where:

y(fit)=A+[(B−A)/(1+((C/x)̂D))].

The activated caspase-3 marker labels cells from early to late stageapoptosis. Concentrations of test compound that cause a 5-fold inductionin the caspase-3 signal (Cal_X5) indicate significant apoptosisinduction. The maximal induction of caspase 3 by compound in comparisonwith DMSO control is reported as Max_Fold_Change.

TABLE 43 Cell lines used in multiplexed cytotoxicity assays Cell LineType Subtype SW-13 Endocrine Adrenal gland NCI-H295R Endocrine Adrenalgland 639-V Bladder Bladder BFTC-905 Bladder Bladder HT1376 BladderBladder SCaBER Bladder Bladder T24 Bladder Bladder 5637 Bladder Bladder647-V Bladder Bladder HT-1197 Bladder Bladder TCCSUP Bladder Bladder J82Bladder Bladder UM-UC-3 Bladder Bladder MDA-MB-436 Breast Breast Hs 578TBreast Breast AU565 Breast Breast BT20 Breast Breast SK-BR-3 BreastBreast BT474 Breast Breast CAMA-1 Breast Breast EFM-19 Breast BreastKPL-1 Breast Breast MDA MB 231 Breast Breast MDA MB 453 Breast BreastMCF7 Breast Breast T47D Breast Breast MDA-MB-415 Breast Breast ZR-75-1Breast Breast BT-549 Breast Breast MDA MB 468 Breast Breast C-33A FemaleGU Cervix C-4 I Female GU Cervix C-4 II Female GU Cervix HeLa Female GUCervix SiHa Female GU Cervix DoTc2 4510 Female GU Cervix HT-3 Female GUCervix LS513 Colon Colon LS411N Colon Colon SNU-C2B Colon Colon LS123Colon Colon MT-3 Colon Colon SW403 Colon Colon RKO-AS45-1 Colon ColonSW480 Colon Colon SW948 Colon Colon Colo 320 HSR Colon Colon HCT-15Colon Colon HCT-116 Colon Colon RKOE6 Colon Colon SW48 Colon Colon SW837Colon Colon SW1463 Colon Colon Colo 320DM Colon Colon HT-29 Colon ColonLS1034 Colon Colon Colo 201 Colon Colon Colo 205 Colon Colon NCI-H747Colon Colon RKO Colon Colon SW1417 Colon Colon DLD-1 Colon ColonNCI-H508 Colon Colon SW620 Colon Colon WiDr Colon Colon HRT-18 ColonColon LS-174T Colon Colon HuTu 80 Duodenum Duodenum Y79 Eye Eye Hs 683Central Nervous System Glioma U-118 MG Central Nervous System GliomaM059J Central Nervous System Glioma PFSK-1 Central Nervous System GliomaSW1783 Central Nervous System Glioma SW1088 Central Nervous SystemGlioma T98G Central Nervous System Glioma CCF-STTG1 Central NervousSystem Glioma A172 Central Nervous System Glioma DBTRG-05MG CentralNervous System Glioma H4 Central Nervous System Glioma SNB-19 CentralNervous System Glioma U-138MG Central Nervous System Glioma U-87 MGCentral Nervous System Glioma DK-MG Central Nervous System Glioma A-253Head and Neck Head and Neck A388 Head and Neck Head and Neck Detroit 562Head and Neck Head and Neck A431 Head and Neck Head and Neck Cal 27 Headand Neck Head and Neck OE19 Head and Neck Head and Neck OE33 Head andNeck Head and Neck SCC-4 Head and Neck Head and Neck FaDu Head and NeckHead and Neck OE21 Head and Neck Head and Neck SCC-25 Head and Neck Headand Neck SCC-9 Head and Neck Head and Neck A-704 Kidney Kidney 769-PKidney Kidney 786-O Kidney Kidney G-402 Kidney Kidney ACHN Kidney KidneyCaki-1 Kidney Kidney Caki-2 Kidney Kidney SK-NEP-1 Kidney Kidney G-401Kidney Kidney A498 Kidney Kidney KG-1 Hematopoietic Leukemia RS4; 11Hematopoietic Leukemia KU812 Hematopoietic Leukemia TF-1 HematopoieticLeukemia MX1 Hematopoietic Leukemia NALM-6 Hematopoietic Leukemia MOLT-3Hematopoietic Leukemia MOLT-16 Hematopoietic Leukemia MEG01Hematopoietic Leukemia MHH-PREB-1 Hematopoietic Leukemia MV-4-11Hematopoietic Leukemia Thp1 Hematopoietic Leukemia BV-173 HematopoieticLeukemia CCRFCEM Hematopoietic Leukemia CML-T1 Hematopoietic LeukemiaHEL-92-1-7 Hematopoietic Leukemia J-RT3-T3-5 Hematopoietic LeukemiaJurkat Hematopoietic Leukemia CEM-C1 Hematopoietic Leukemia EM-2Hematopoietic Leukemia K562 Hematopoietic Leukemia HuCCT1 Liver LiverHLE Liver Liver HUH-6 Clone 5 Liver Liver HepG2 Liver Liver HLF LiverLiver OCUG-1 Liver Liver SNU-423 Liver Liver Hs 611.T HematopoieticLymphoma EB2 Hematopoietic Lymphoma GA-10 Hematopoietic Lymphoma H9Hematopoietic Lymphoma JeKo-1 Hematopoietic Lymphoma SU-DHL-8Hematopoietic Lymphoma SUP-T1 Hematopoietic Lymphoma TUR HematopoieticLymphoma Hs 445 Hematopoietic Lymphoma BCP-1 Hematopoietic Lymphoma CA46Hematopoietic Lymphoma Jiyoye Hematopoietic Lymphoma MC116 HematopoieticLymphoma NAMALWA Hematopoietic Lymphoma REC-1 Hematopoietic LymphomaSU-DHL-4 Hematopoietic Lymphoma SU-DHL-5 Hematopoietic LymphomaSU-DHL-10 Hematopoietic Lymphoma DB Hematopoietic Lymphoma DOHH-2Hematopoietic Lymphoma HT Hematopoietic Lymphoma RPMI 6666 HematopoieticLymphoma Raji Hematopoietic Lymphoma SR Hematopoietic Lymphoma ST486Hematopoietic Lymphoma BC-1 Hematopoietic Lymphoma Daudi HematopoieticLymphoma L-428 Hematopoietic Lymphoma EB-3 Hematopoietic Lymphoma Ramos(RA 1) Hematopoietic Lymphoma CRO-AP2 Hematopoietic Lymphoma D341 MedCentral Nervous System Medulloblastoma D283 Med Central Nervous SystemMedulloblastoma Daoy Central Nervous System Medulloblastoma Hs 852.TSkin (Melanoma) Melanoma WM-266-4 Skin (Melanoma) Melanoma Hs 934.T Skin(Melanoma) Melanoma A2058 Skin (Melanoma) Melanoma G-361 Skin (Melanoma)Melanoma Hs 688(A).T Skin (Melanoma) Melanoma Hs 936.T(C1) Skin(Melanoma) Melanoma Hs 895.T Skin (Melanoma) Melanoma A7 Skin (Melanoma)Melanoma C32 Skin (Melanoma) Melanoma CHL-1 Skin (Melanoma) MelanomaSK-MEL-28 Skin (Melanoma) Melanoma SH-4 Skin (Melanoma) MelanomaRPMI-7951 Skin (Melanoma) Melanoma MALME3M Skin (Melanoma) Melanoma MeWoSkin (Melanoma) Melanoma SK-MEL-1 Skin (Melanoma) Melanoma SK-MEL-3 Skin(Melanoma) Melanoma C32TG Skin (Melanoma) Melanoma Hs 294T Skin(Melanoma) Melanoma Hs 695T Skin (Melanoma) Melanoma A101D Skin(Melanoma) Melanoma A375 Skin (Melanoma) Melanoma COLO 829 Skin(Melanoma) Melanoma HMCB Skin (Melanoma) Melanoma IM-9 HematopoieticMyeloma SKO-007 Hematopoietic Myeloma U26661 Hematopoietic Myeloma RPMI8226 Hematopoietic Myeloma ARH-77 Hematopoietic Myeloma BE(2)C CentralNervous System Neuroblastoma SK-N-FI Central Nervous SystemNeuroblastoma CHP-212 Central Nervous System Neuroblastoma SK-N-ASCentral Nervous System Neuroblastoma MC-IXC Central Nervous SystemNeuroblastoma SK-N-DZ Central Nervous System Neuroblastoma Hs 229.T LungNSCLC NCI-H661 Lung NSCLC A427 Lung NSCLC Calu6 Lung NSCLC NCI-H460 LungNSCLC NCI-H520 Lung NSCLC NCI-H596 Lung NSCLC NCIH441 Lung NSCLC A549Lung NSCLC ChaGoK1 Lung NSCLC Calu1 Lung NSCLC COR-L23 Lung NSCLC SKMES1Lung NSCLC NCI-H292 Lung NSCLC COR-L105 Lung NSCLC G-292, clone A141B1Soft Tissue Osteosarcoma Hs 888.Sk Soft Tissue Osteosarcoma HOS SoftTissue Osteosarcoma MG-63 Soft Tissue Osteosarcoma SJSA1 Soft TissueOsteosarcoma SW1353 Soft Tissue Osteosarcoma SaO52 Soft TissueOsteosarcoma U2OS Soft Tissue Osteosarcoma KHOS-24OS Soft TissueOsteosarcoma ME-180 Female GU Ovary PA-1 Female GU Ovary Ca Ski FemaleGU Ovary M5751 Female GU Ovary CaOV3 Female GU Ovary OVCAR3 Female GUOvary SKOV3 Female GU Ovary PSN-1 Pancreas Pancreas AsPC-1 PancreasPancreas PANC-1 Pancreas Pancreas Hs 766T Pancreas Pancreas Mia PaCa-2Pancreas Pancreas SU.86.86 Pancreas Pancreas YAPC Pancreas PancreasBxPC-3 Pancreas Pancreas CFPAC-1 Pancreas Pancreas Capan-1 PancreasPancreas Capan-2 Pancreas Pancreas HPAF-II Pancreas Pancreas HuP-T4Pancreas Pancreas BeWo Placenta Placenta JAR Placenta Placenta JEG-3Placenta Placenta 22Ry1 Prostate Prostate DU145 Prostate Prostate PC-3Prostate Prostate LNCaP Prostate Prostate BM-1604 Prostate Prostate BPH1Prostate Prostate Hs 729 Soft Tissue Sarcoma VA-ES-BJ Soft TissueSarcoma Hs 821.T Soft Tissue Sarcoma TE 125.T Soft Tissue Sarcoma RDSoft Tissue Sarcoma SK-UT-1 Soft Tissue Sarcoma A-673 Soft TissueSarcoma SW684 Soft Tissue Sarcoma A204 Soft Tissue Sarcoma SW872 SoftTissue Sarcoma SW982 Soft Tissue Sarcoma HT-1080 Soft Tissue SarcomaMES-SA Soft Tissue Sarcoma SJRH30 Soft Tissue Sarcoma SK-LMS-1 SoftTissue Sarcoma TE 381.T Soft Tissue Sarcoma NCI-H510A Lung SCLC NCIH446Lung SCLC SHP-77 Lung SCLC DMS114 Lung SCLC SW900 Lung SCLC DMS53 LungSCLC NCI-H69 Lung SCLC DMS273 Lung SCLC SK-PN-DW Stomach Stomach AGSStomach Stomach HS 746T Stomach Stomach SNU-1 Stomach Stomach KATO IIIStomach Stomach SNU-16 Stomach Stomach SNU-5 Stomach Stomach NTERA-2cl.D1 Testis Testis TT Endocrine Thyroid BHT-101 Endocrine ThyroidCAL-62 Endocrine Thyroid CGTH-W-1 Endocrine Thyroid SW579 EndocrineThyroid HEC-1-A Female GU Uterus RL95-2 Female GU Uterus KLE Female GUUterus AN3 CA Female GU Uterus SW962 Female GU Vulva SW954 Female GUVulva

The solid forms of Compound 1 described herein show or will be shown tohave anti-proliferative activity in a variety of cancer cell lines.Anti-proliferative activity in these cancer cell lines indicates thatthe Aminopurine compounds may be useful in the treatment of cancers,including solid tumors, as exemplified by melanoma, colorectal cancer,stomach cancer, head and neck cancer, thyroid cancer, bladder cancer,CNS cancer, lung cancer, pancreatic cancer, and soft tissue cancer.

In another embodiment, solid forms of Compound 1 described herein showor will be shown to induce apoptosis in a variety of cancer cell lines.Induction of apoptosis indicates that the solid forms of Compound 1described herein may be useful in the treatment of cancers, includingsolid tumors, as exemplified by bladder cancer, breast cancer, CNScancer (including neuroblastoma and glioma), colon cancer,gastrointestinal cancer (for example, stomach cancer or colon cancer),endocrine cancer (for example, thyroid cancer or adrenal gland cancer),female genitoureal cancer (for example, cervix cancer or ovary clearcell cancer, vulva cancer, uterus cancer, or ovary cancer), head andneck cancer, hematopoietic cancer (for example, leukemia or myeloma),kidney cancer, liver cancer, lung (for example, NSCLC or SCLC),melanoma, pancreas cancer, prostate cancer, or soft tissue cancer (forexample, sarcoma or osteosarcoma).

In another embodiment, solid forms of Compound 1 described herein showor will be shown to cause G1/S arrest in a variety of cancer cell lines.Causing G1/S arrest in these cancer cell lines indicates that thecompounds may be useful in the treatment of cancers, including solidtumors, as exemplified by bladder cancer, breast cancer, CNS cancer (forexample, glioma or neuroblastoma), colon cancer, gastrointestinal cancer(for example, stomach cancer), endocrine cancer (for example, thyroidcancer or adrenal gland cancer), female genitoureal cancer (for example,uterus cancer, cervix cancer, ovary clear cell cancer, or vulva cancer),head and neck cancer, hematopoietic cancer (for example, leukemia ormyeloma), kidney cancer, liver cancer, lung cancer (for example, NSCLCor SCLC), melanoma, pancreas cancer, prostate cancer, or soft tissuecancer (sarcoma or osteosarcoma).

Multiplexed Cytotoxicity Assay.

In another experiment, cells were grown in RPMI1640, 10% FBS, 2 mML-alanyl-L-Glutamine, 1 mM Na pyruvate or a special medium in ahumidified atmosphere of 5% CO₂ at 37° C. Cells were seeded into384-well plates and incubated in a humidified atmosphere of 5% CO₂ at37° C. Compounds were added 24 h post cell seeding. At the same time, atime zero untreated cell plate was generated. After a 72 hour incubationperiod, cells were fixed and stained with fluorescently labeledantibodies and nuclear dye to allow visualization of nuclei, apoptoticcells and mitotic cells. Apoptotic cells were detected using ananti-active caspase-3 antibody. Mitotic cells were detected using ananti phospho-histone-3 antibody. Compounds were serially diluted3.16-fold and assayed over 10 concentrations in a final assayconcentration of 0.1% DMSO from the highest test concentration of 10 μM.Automated fluorescence microscopy was carried out using a MolecularDevices ImageXpress Micro XL high-content imager, and images werecollected with a 4× objective.

Data Analysis.

Sixteen-bit TIFF images were acquired and analyzed with MetaXpress5.1.0.41 software. Cell proliferation was measured by the signalintensity of the incorporated nuclear dye. The cell proliferation assayoutput was referred to as the relative cell count. To determine the cellproliferation end point, the cell proliferation data output wastransformed to percentage of control (POC) using the following formula:

POC=relative cell count(compound wells)/relative cell count(vehiclewells)×100

Relative cell count IC₅₀ was the test compound concentration at 50% ofmaximal possible response relative to the DMSO control. GI₅₀ refers tothe concentration needed to reduce the observed growth by half. Thiscorresponds to the concentration that inhibits the growth to the levelmidway between growth in untreated cells and the number of cells seededin the well (Time zero value). The IC₅₀ values were calculated usingnonlinear regression to fit data to a sigmoidal 4 point, 4 parameterOne-Site dose response model, where:

y(fit)=A+[(B−A)/(1+((C/x)̂D))].

The activated caspase-3 marker labels cells from early to late stageapoptosis. Concentrations of test compound that cause a 2-fold (Cal-X2)or 5-fold induction in the caspase-3 signal (Cal_X5) indicatedsignificant apoptosis induction. The maximal induction of caspase 3 bycompound in comparison with DMSO control was reported asMax_Fold_Change.

TABLE 44 Results of Cytotoxicity Assays Max GI50 IC50 CalX2 CalX5 foldCell line Tumor Type Subtype (μM) (μM) (μM) (μM) change NCIH295REndocrine Adrenal gland 10 10 10 10 1.74 SW13 Endocrine Adrenal gland0.0711 0.135 0.04 0.535 8.74 5637 Bladder Bladder 6.85 9.77 10 10 2.12639V Bladder Bladder 0.184 0.206 0.0841 0.465 6.33 647V Bladder Bladder6.93 7.82 2.7413 4.45 19.29 BFTC905 Bladder Bladder 0.0515 0.0546 0.01790.0414 45.3 HT1197 Bladder Bladder 0.444 10 0.1601 10 4.16 HT1376Bladder Bladder 0.792 3.48 0.0524 0.167 10.87 J82 Bladder Bladder 10 102.4365 10 3.17 SCABER Bladder Bladder 0.0665 0.0772 0.0086 0.0506 29.47T24 Bladder Bladder 0.233 0.274 4.5443 10 2.61 TCCSUP Bladder Bladder2.21 6.59 5.6435 10 3.67 UMUC3 Bladder Bladder 0.149 0.201 2.7934 5.766.56 AU565 Breast Breast 8.15 8.77 3.8749 7.14 14.18 BT20 Breast Breast8.36 10 10 10 1.81 BT474 Breast Breast 10 10 10 10 0.94 BT549 BreastBreast 10 10 5.4537 10 3.14 CAMA1 Breast Breast 0.298 2.24 6.4981 102.85 EFM19 Breast Breast 4.2 10 10 10 2.1 HS578T Breast Breast 0.1530.837 2.6723 6.58 5.94 KPL1 Breast Breast 10 10 0.0481 10 2.63 MCF7Breast Breast 0.636 3.47 6.5592 9.74 5.78 MDAMB231 Breast Breast 0.03390.0624 0.0242 0.257 5.94 MDAMB415 Breast Breast 0.729 10 10 10 1.85MDAMB436 Breast Breast 0.262 10 5.118 10 4.25 MDAMB453 Breast Breast0.656 2.82 10 10 1.07 MDAMB468 Breast Breast 0.0363 0.0721 0.0969 103.81 MT3 Breast Breast 0.674 1.08 7.6544 10 2.81 SKBR3 Breast Breast6.81 8.45 3.2211 6.2 12.79 T47D Breast Breast 10 10 10 10 2 ZR751 BreastBreast 0.0943 7.7 5.9055 6.44 7.36 A431 Skin Carcinoma 0.228 0.3110.0801 1.76 5.11 C33A Female GU Cervix 0.191 0.407 3.6798 5.39 9.45 C4IFemale GU Cervix 10 10 5.7177 7.94 7.38 C4II Female GU Cervix 10 100.044 10 3.7 DOTC24510 Female GU Cervix 0.04 0.132 0.0268 10 5.03 HELAFemale GU Cervix 6.75 8.71 7.0794 10 3.65 HT3 Female GU Cervix 0.8563.21 0.2906 3.74 7.49 SIHA Female GU Cervix 10 10 7.6882 8.82 5.49COLO201 Colon Colon 0.0128 0.0172 0.0225 0.267 6.09 COLO205 Colon Colon0.0095 0.0117 0.0102 0.0248 9.86 COLO320DM Colon Colon 9.11 10 6.18629.53 5.28 COLO320HSR Colon Colon 4.19 4.44 2.0186 3.53 49.73 DLD1 ColonColon 0.162 0.197 0.0474 0.104 21.95 HCT116 Colon Colon 0.0194 0.02040.0196 0.0448 45.43 HCT15 Colon Colon 1.97 2.23 5.1211 7.03 7.97 HRT18Colon Colon 0.0775 0.0819 0.0657 0.147 11.1 HT29 Colon Colon 0.01290.0167 0.0092 0.0318 61.59 LS1034 Colon Colon 0.224 0.676 1.4781 10 2.52LS123 Colon Colon 0.061 0.188 0.0766 10 4.74 LS174T Colon Colon 0.1940.259 0.2846 0.412 5.63 LS411N Colon Colon 0.0358 0.053 0.0575 10 5.58LS513 Colon Colon 0.0353 0.0386 0.0233 0.0356 64.31 NCIH508 Colon Colon0.0288 0.0481 0.0778 1.25 5.37 NCIH747 Colon Colon 0.012 0.0445 0.02260.0756 8.21 RKO Colon Colon 0.0353 0.0405 0.0407 0.378 11.14 RKOAS451Colon Colon 0.0405 0.0449 0.1873 1.16 10.06 RKOE6 Colon Colon 0.07530.107 1.6988 3.6 29.26 SNUC2B Colon Colon 0.0544 0.722 10 10 1.67 SW1417Colon Colon 0.0088 0.0351 0.0221 0.0693 6.76 SW1463 Colon Colon 0.1350.181 2.4138 10 2.82 SW403 Colon Colon 0.0476 0.173 0.1084 10 4.02 SW48Colon Colon 0.0018 0.0031 0.0047 0.0266 13.66 SW480 Colon Colon 0.01840.0311 0.0638 0.248 6.26 SW620 Colon Colon 0.0492 0.0798 1.4774 3.8814.66 SW837 Colon Colon 0.172 0.348 0.325 10 4.34 SW948 Colon Colon0.195 0.327 10 10 1.57 WIDR Colon Colon 0.0104 0.0133 0.0085 0.021 79.03HUTU80 Duodenum Duodenum 0.057 0.0695 0.0161 0.354 9.27 Y79 Eye- Eye 1010 7.8739 10 2.58 retinoblastoma A172 CNS Glioma 0.0649 0.139 0.11742.36 5.95 CCFSTTG1 CNS Glioma 10 10 10 10 1.03 DBTRG05MG CNS Glioma0.0432 0.0984 0.1963 10 3.94 DKMG CNS Glioma 0.0207 0.126 0.0463 0.1610.86 H4 CNS Glioma 0.758 0.943 1.7285 3.78 14.47 HS683 CNS Glioma 0.1480.305 10 10 2.54 M059J CNS Glioma 0.612 3.31 4.9633 10 2.8 PFSK1 CNSGlioma 0.0234 10 10 10 1.06 SNB19 CNS Glioma 0.163 0.244 0.4478 10 3.29SW1088 CNS Glioma 3.35 5.98 5.2615 7.5 9.59 SW1783 CNS Glioma 5.92 9.859.0994 10 2.49 T98G CNS Glioma 10 10 5.4225 10 3.16 U118MG CNS Glioma0.175 10 10 10 1.92 U138MG CNS Glioma 0.053 10 0.1598 0.417 8.01 U87MGCNS Glioma 0.0692 0.101 9.3615 10 2.14 A253 Head and Neck Head and Neck0.171 10 8.7811 10 2.85 A388 Head and Neck Head and Neck 0.422 1.120.0902 3.52 6.48 CAL27 Head and Neck Head and Neck 0.0592 0.0661 0.08770.46 7.98 DETROIT562 Head and Neck Head and Neck 0.347 10 4.9484 7.166.02 FADU Head and Neck Head and Neck 0.435 0.787 4.0608 5.64 8.64 SCC25Head and Neck Head and Neck 0.0439 0.051 0.1187 0.304 6.72 SCC4 Head andNeck Head and Neck 0.0512 0.108 0.0317 0.065 7.38 SCC9 Head and NeckHead and Neck 0.117 0.28 0.6679 3.86 9.58 769P Kidney Kidney 0.194 0.2550.2023 5.11 5.67 786O Kidney Kidney 2.04 6.92 10 10 0.83 A498 KidneyKidney 0.522 0.808 0.5562 10 4.72 A704 Kidney Kidney 10 10 10 10 0.96ACHN Kidney Kidney 0.306 0.55 0.78 10 2.97 CAKI1 Kidney Kidney 0.09140.151 0.2015 10 4.12 CAKI2 Kidney Kidney 0.139 0.193 0.1631 0.449 6.26G401 Kidney Kidney 0.0774 0.086 0.0717 0.179 30.87 G402 Kidney Kidney0.0504 0.0925 0.0162 0.637 7.34 SKNEP1 Kidney Kidney 10 10 10 10 1.15BV173 Hematopoietic Leukemia 1.1 10 0.4959 10 2.91 and lymphoid CCRFCEMHematopoietic Leukemia 5.03 6.05 3.4279 6.95 12.74 and lymphoid CEMC1Hematopoietic Leukemia 10 10 4.1828 5.22 11.27 and lymphoid CMLT1Hematopoietic Leukemia 0.149 10 0.0948 10 4.85 and lymphoid EM2Hematopoietic Leukemia 0.0481 0.0936 10 10 1.55 and lymphoid HEL9217Hematopoietic Leukemia 4.62 8.23 3.2991 6.57 6.23 and lymphoid JRT3T35Hematopoietic Leukemia 3.58 4.78 2.6364 3.8 14.26 and lymphoid JURKATHematopoietic Leukemia 3.34 3.73 1.6173 3.28 14.48 and lymphoid K562Hematopoietic Leukemia 10 10 2.9298 4.86 51.89 and lymphoid KG1Hematopoietic Leukemia 0.0017 0.0325 2.5811 10 2.5 and lymphoid KU812Hematopoietic Leukemia 0.003 0.0159 0.03 8.02 5.63 and lymphoid MEG01Hematopoietic Leukemia 0.0818 0.221 0.5718 10 2.77 and lymphoid MHHPREB1Hematopoietic Leukemia 6.69 6.97 5.1142 7.66 11.43 and lymphoid MOLT16Hematopoietic Leukemia 2.88 3.35 2.4102 4.97 8.06 and lymphoid MOLT3Hematopoietic Leukemia 0.946 3.03 5.88 10 3.63 and lymphoid MV411Hematopoietic Leukemia 0.107 0.184 0.0933 1.15 8.12 and lymphoid MX1Hematopoietic Leukemia 0.0401 0.0619 1.1016 10 3.78 and lymphoid NALM6Hematopoietic Leukemia 10 10 0.1241 10 5 and lymphoid RS411Hematopoietic Leukemia 0.359 2.96 3.8025 8.4 5.83 and lymphoid TF1Hematopoietic Leukemia 0.0015 0.0095 0.006 0.0296 16.1 and lymphoid THP1Hematopoietic Leukemia 0.0251 0.0495 0.132 3.9 6.3 and lymphoid HEPG2Liver Liver 0.0224 0.0643 0.0041 0.0108 62.47 HLE Liver Liver 0.683 1.040.8174 10 2.5 HLF Liver Liver 4.76 6.47 10 10 1.95 HUCCT1 Liver Liver0.0537 0.0633 0.0222 0.0406 11.54 HUH6CLONE5 Liver Liver 0.145 0.3540.0631 0.302 8.25 OCUG1 Liver Liver 0.464 1.29 0.0848 0.49 5.49 SNU423Liver Liver 0.192 0.276 0.0909 1.65 7.32 BC1 Hematopoietic Lymphoma 1010 5.1005 6.54 8.72 and lymphoid BCP1 Hematopoietic Lymphoma 0.02050.0797 4.8663 7.36 6.56 and lymphoid CA46 Hematopoietic Lymphoma 0.01460.0213 3.2395 8.08 9.46 and lymphoid CROAP2 Hematopoietic Lymphoma 0.9962.58 2.9603 4.2 50.79 and lymphoid DAUDI Hematopoietic Lymphoma 0.017710 3.9392 5.33 10.08 and lymphoid DB Hematopoietic Lymphoma 0.0131 106.1153 6.5 7.11 and lymphoid DOHH2 Hematopoietic Lymphoma 5.54 5.792.4833 3.99 20.41 and lymphoid EB2 Hematopoietic Lymphoma 0.389 0.555.7381 10 4.16 and lymphoid EB3 Hematopoietic Lymphoma 1.63 2.15 6.14697.66 5.5 and lymphoid GA10 Hematopoietic Lymphoma 0.0468 0.0567 0.64771.94 6.49 and lymphoid H9 Hematopoietic Lymphoma 0.0232 0.039 0.0222 0.47.33 and lymphoid HS445 Hematopoietic Lymphoma 0.0143 0.0377 4.9128 7.75.65 and lymphoid HS611T Hematopoietic Lymphoma 0.0106 0.0123 2.8507 103.84 and lymphoid HT Hematopoietic Lymphoma 8.3 10 8.6354 10 2.44 andlymphoid JEKO1 Hematopoietic Lymphoma 0.461 0.83 4.3369 10 3.11 andlymphoid JIYOYE Hematopoietic Lymphoma 0.0814 0.21 4.4004 5.35 11.1 andlymphoid L428 Hematopoietic Lymphoma 1.63 3.46 4.2384 5.88 7.51 andlymphoid MC116 Hematopoietic Lymphoma 6.02 6.49 2.8763 5.18 9.46 andlymphoid NAMALWA Hematopoietic Lymphoma 0.0181 0.0239 5.9431 10 2.68 andlymphoid RAJI Hematopoietic Lymphoma 0.179 10 2.5564 4.07 24.81 andlymphoid RAMOSRA1 Hematopoietic Lymphoma 3.66 3.84 4.5496 7.39 25.1 andlymphoid REC1 Hematopoietic Lymphoma 0.0053 0.193 10 10 1.86 andlymphoid RPMI6666 Hematopoietic Lymphoma 0.0801 0.37 3.0419 4.37 26.35and lymphoid SR Hematopoietic Lymphoma 1.42 1.84 1.2842 3.07 33.52 andlymphoid ST486 Hematopoietic Lymphoma 5.02 6.14 4.2422 6.11 10.85 andlymphoid SUDHL10 Hematopoietic Lymphoma 1.23 1.4 3.611 4.87 11.63 andlymphoid SUDHL4 Hematopoietic Lymphoma 0.168 0.332 2.5668 4.83 10.75 andlymphoid SUDHL5 Hematopoietic Lymphoma 0.0011 0.0013 1.6359 4.54 10.37and lymphoid SUDHL8 Hematopoietic Lymphoma 0.0193 0.0406 1.2344 4.1910.79 and lymphoid SUPT1 Hematopoietic Lymphoma 0.0196 0.0466 4.54769.21 5.76 and lymphoid TUR Hematopoietic Lymphoma 0.0415 0.0539 0.69843.45 17.35 and lymphoid D283MED CNS Medulloblastoma 2.56 7.55 8.3456 102.23 D341MED CNS Medulloblastoma 10 0.0219 7.7855 10 2.14 DAOY CNSMedulloblastoma 0.749 1.09 3.2773 5.22 16.68 A101D Skin Melanoma 0.04240.0815 0.4207 3.71 7.93 A2058 Skin Melanoma 0.212 0.288 0.065 0.20411.68 A375 Skin Melanoma 0.0065 0.0072 0.0673 0.0827 103.79 A7 SkinMelanoma 1.72 7.27 5.0814 9.4 5.51 C32 Skin Melanoma 0.0289 0.111 0.04510.0778 110.9 C32TG Skin Melanoma 0.0408 0.109 0.0608 0.117 42.82 CHL1Skin Melanoma 0.103 0.117 1.2376 10 3.46 COLO829 Skin Melanoma 0.01210.0343 0.0421 0.125 24.28 G361 Skin Melanoma 0.102 0.15 0.0428 0.1224.48 HMCB Skin Melanoma 0.0724 0.113 10 10 1.8 HS294T Skin Melanoma0.0507 0.0706 0.154 2.15 5.74 HS688AT Skin Melanoma 0.0822 10 10 10 1.61HS695T Skin Melanoma 0.0363 0.16 0.0253 0.0727 22.05 HS852T SkinMelanoma 0.0564 0.715 0.05 0.234 6.51 HS895T Skin Melanoma 10 10 10 101.52 HS934T Skin Melanoma 0.0052 10 0.3638 1.4 5.1 HS936TC1 SkinMelanoma 0.0184 0.0258 0.0084 0.0224 134.98 MALME3M Skin Melanoma 0.00340.012 0.0025 0.0045 102.73 MEWO Skin Melanoma 0.102 0.159 0.167 0.37314.34 RPMI7951 Skin Melanoma 0.0716 0.0945 0.1237 1.29 28.15 SH4 SkinMelanoma 0.0208 0.029 0.0157 0.0382 66.44 SKMEL1 Skin Melanoma 0.0010.0291 0.1019 0.194 7.63 SKMEL28 Skin Melanoma 0.0279 0.0571 0.29070.344 16.64 SKMEL3 Skin Melanoma 0.0284 0.0625 10 10 1.74 WM2664 SkinMelanoma 0.012 0.0354 0.0023 0.0151 83.29 ARH77 Hematopoietic Myeloma 1010 10 10 1.86 and lymphoid IM9 Hematopoietic Myeloma 0.0911 0.143 0.04310 4.85 and lymphoid RPMI8226 Hematopoietic Myeloma 1.09 2.48 3.31035.34 8.35 and lymphoid SKO007 Hematopoietic Myeloma 0.0274 0.482 0.17582.79 7.24 and lymphoid U266B1 Hematopoietic Myeloma 0.0133 0.109 0.049310 4.36 and lymphoid BE2C CNS Neuroblastoma 0.146 0.21 0.1223 10 5.47CHP212 CNS Neuroblastoma 0.0066 0.0165 0.019 0.341 5.97 MCIXC CNSNeuroblastoma 2.04 2.33 1.9309 4.62 5.15 SKNAS CNS Neuroblastoma 0.04890.132 0.0675 0.227 8.86 SKNDZ CNS Neuroblastoma 7.4 10 10 10 1.23 SKNFICNS Neuroblastoma 0.0151 0.135 0.0897 10 3.51 A427 Lung NSCLC 0.04750.0763 0.0018 10 3.34 A549 Lung NSCLC 0.102 0.128 0.0297 0.0946 13.03CALU1 Lung NSCLC 0.0967 0.149 0.2575 10 3.73 CALU6 Lung NSCLC 0.04630.083 0.11 10 4.86 CHAGOK1 Lung NSCLC 10 10 10 10 1.23 CORL105 LungNSCLC 0.0165 0.0414 0.0583 0.571 6.55 CORL23 Lung NSCLC 0.0238 0.02830.0176 0.0569 12.96 HS229T Lung NSCLC 0.415 10 0.8448 7.22 5.29 NCIH292Lung NSCLC 0.278 0.686 2.4602 4.85 10.26 NCIH441 Lung NSCLC 0.271 1.257.7406 10 4.32 NCIH460 Lung NSCLC 10 10 10 10 0.98 NCIH520 Lung NSCLC0.991 2.13 3.637 5.03 13.95 NCIH596 Lung NSCLC 2.75 10 10 10 1.11NCIH661 Lung NSCLC 1.44 2.64 0.0833 10 4.58 SKMES1 Lung NSCLC 0.1030.122 0.0384 0.212 27.03 OE19 Head and Neck Esophageal 0.34 10 10 101.79 OE21 Head and Neck Esophageal 0.0939 0.124 0.0221 0.948 5.91 OE33Head and Neck Esophageal 0.063 0.0969 0.0317 0.495 5.9 G292CLONEA141B1Soft Tissue Osteosarcoma 0.0272 0.0493 0.0401 0.211 7.61 HOS Soft TissueOsteosarcoma 2.57 3.69 6.2324 8.81 7.12 HS888SK Soft Tissue Osteosarcoma0.111 10 0.1023 0.175 15.7 KHOS240S Soft Tissue Osteosarcoma 10 104.3797 4.93 18.16 MG63 Soft Tissue Osteosarcoma 0.108 0.115 4.1626 5.7117.21 SAOS2 Soft Tissue Osteosarcoma 3.57 6.88 3.2386 5.98 6.35 SJSA1Soft Tissue Osteosarcoma 1.16 2.46 2.9744 6.21 62.65 SW1353 Soft TissueOsteosarcoma 0.184 0.292 0.404 10 4.79 U2OS Soft Tissue Osteosarcoma0.23 0.373 0.0332 0.0801 20.57 CAOV3 Female GU Ovary 0.429 10 2.0076 102.95 CASKI Female GU Ovary 6.76 10 0.9719 10 2.61 ME180 Female GU Ovary10 10 5.1674 6.32 12.19 MS751 Female GU Ovary 6.91 9.51 5.4363 10 3.62OVCAR3 Female GU Ovary 10 10 10 10 1.19 PA1 Female GU Ovary 0.471 2.623.6547 5.1 11.55 SKOV3 Female GU Ovary 0.547 10 0.2939 10 2.65 ASPC1Pancreas Pancreas 0.0308 10 0.0471 10 4.08 BXPC3 Pancreas Pancreas0.0369 0.0455 0.025 10 4.98 CAPAN1 Pancreas Pancreas 0.105 10 10 10 1.97CAPAN2 Pancreas Pancreas 0.136 0.291 10 0.209 6.62 CFPAC1 PancreasPancreas 10 10 10 10 1.46 HPAFII Pancreas Pancreas 0.013 0.0175 0.00340.0093 52.77 HS766T Pancreas Pancreas 0.0343 0.0793 0.0646 0.632 6.36HUPT4 Pancreas Pancreas 0.0434 0.0505 0.0998 10 5.3 MIAPACA2 PancreasPancreas 0.0357 0.0396 0.0387 0.578 15.16 PANC1 Pancreas Pancreas 0.04160.08 0.0227 0.173 10.76 PSN1 Pancreas Pancreas 0.0083 0.0092 0.0360.0701 8.75 SU8686 Pancreas Pancreas 0.0635 0.132 10 10 2.09 YAPCPancreas Pancreas 0.183 0.67 10 10 1.59 BEWO Female GU Placenta 5.165.69 3.9778 6.42 10.1 JAR Female GU Placenta 3.17 3.21 1.0062 2.99102.66 JEG3 Female GU Placenta 6.34 7.75 5.8823 7.95 6.39 22RV1 ProstateProstate 2.66 5.58 3.0283 4.45 18.69 BM1604 Prostate Prostate 0.1410.401 10 10 1.8 BPH1 Prostate Prostate 0.0578 0.0675 0.0577 0.116 35.09DU145 Prostate Prostate 0.0738 0.0965 5.1233 8.37 6.15 LNCAP ProstateProstate 2.43 5.07 4.1807 10 3.85 PC3 Prostate Prostate 7.82 8.54 10 103.64 A204 Soft Tissue Sarcoma 10 10 0.2906 10 3.48 A673 Soft TissueSarcoma 3.75 3.87 3.411 4.59 27.78 HS729 Soft Tissue Sarcoma 0.54 10 1010 1.87 HS821T Soft Tissue Sarcoma 0.169 10 10 10 1.53 HT1080 SoftTissue Sarcoma 0.0648 0.0727 0.0509 0.107 63.63 MESSA Soft TissueSarcoma 0.81 1.1 4.196 5.47 8.03 RD Soft Tissue Sarcoma 0.0367 0.04430.0297 0.0581 14.86 SJRH30 Soft Tissue Sarcoma 0.219 1.47 0.039 10 5.61SKLMS1 Soft Tissue Sarcoma 0.146 0.166 0.1405 0.876 12.5 SKUT1 SoftTissue Sarcoma 10 10 6.5345 10 4.63 SW684 Soft Tissue Sarcoma 0.08690.37 0.256 0.308 16.88 SW872 Soft Tissue Sarcoma 0.105 0.136 0.05380.434 9.48 SW982 Soft Tissue Sarcoma 0.0156 0.0614 10 10 1.94 TE125TSoft Tissue Sarcoma 1.09 10 3.9673 10 2.5 TE381T Soft Tissue Sarcoma0.0076 0.0128 0.0048 0.0143 15.88 VAESBJ Soft Tissue Sarcoma 0.336 0.583.1752 10 3.26 DMS114 Lung SCLC 0.0688 0.6 0.9142 10 3.38 DMS273 LungSCLC 5.96 6.79 6.5676 8.53 6.76 DMS53 Lung SCLC 0.998 10 0.0661 1.4 7.01NCIH446 Lung SCLC 0.327 10 10 10 1.63 NCIH510A Lung SCLC 3.7 6.61 3.85178.62 6.44 NCIH69 Lung SCLC 5 10 10 10 1.7 SHP77 Lung SCLC 4.79 5.826.8591 10 3.64 SW900 Lung SCLC 0.0216 0.0399 0.0162 0.0849 10.26 AGSStomach Stomach 0.0086 0.0098 0.0075 0.0131 31.12 HS746T Stomach Stomach0.0396 0.122 0.0471 10 4.41 KATOIII Stomach Stomach 0.0612 0.0787 0.01370.123 29.59 SKPNDW Stomach Stomach 3.6 10 7.8388 10 2.58 SNU1 StomachStomach 0.0355 0.0631 0.041 2.57 5.5 SNU16 Stomach Stomach 10 10 3.29685.11 10.66 SNU5 Stomach Stomach 0.0368 0.0943 0.1664 10 3.21 NTERA2CLD1Testis Testis 0.044 0.0507 0.0707 0.0957 9.95 BHT101 Endocrine Thyroid0.0376 0.0412 0.0438 0.0864 22.52 CAL62 Endocrine Thyroid 0.0836 0.09360.0795 0.129 6.49 CGTHW1 Endocrine Thyroid 0.0547 0.0605 0.065 0.10391.55 SW579 Endocrine Thyroid 0.0477 0.0708 0.1374 0.256 51.22 TTEndocrine Thyroid 0.0863 10 0.5946 10 2.79 AN3CA Female GU Uterus 0.7137.03 8.777 10 2.75 HEC1A Female GU Uterus 1.8 2.81 1.6552 3.7 30.12 KLEFemale GU Uterus 10 10 10 10 1.37 RL952 Female GU Uterus 0.009 0.05990.1762 10 4.12 SW954 Female GU Vulva 0.114 0.142 0.1749 0.521 9.14 SW962Female GU Vulva 0.0828 0.232 0.0686 10 3.39

Effect on HCC Proliferation.

HCC cell lines were treated with DMSO or increasing concentrations ofCompound 1 for 72 h. Specifically, Compound 1 at various concentrationsin dimethyl sulfoxide (DMSO) was spotted via an acoustic dispenser (EDCATS-100) into an empty 384-well plate. Compound 1 was spotted in a10-point serial dilution fashion (3-fold dilution) in duplicate withinthe plate. Replicates of plates spotted with Compound 1 were made foruse with different cell lines. After compound plate replication, allplates were sealed (Agilent ThermoLoc) and stored at −20° C. for up to 1month. When ready for testing, plates were removed from the freezer,thawed, and unsealed just prior to the addition of the test cells.

Prior to testing, cells were grown and expanded in culture flasks toprovide sufficient amounts of starting material. Cells were then dilutedto the appropriate densities and added directly to the compound-spotted384-well plates. Cells were allowed to grow for 72 h at 37° C./5% CO₂.At the time when compound was added (t₀), initial cell number wasassessed via a viability assay (Cell Titer-Glo) by quantifying the levelof luminescence generated by ATP present in viable cells. After 72 h,cell viability of compound-treated cells was assessed via Cell Titer-Gloand luminescence measurement. The apoptotic response to Compound 1 wasassessed by quantifying the activities of caspase 3 and caspase 7(Caspase 3/7-Glo) in treated cells and DMSO control cells.

Determination of G1₅₀ and IC₅₀ Values.

A Four Parameter Logistic Model (Sigmoidal Dose-Response Model) was usedto determine the compound's GI₅₀ value.

y=(A+((B−A)/(1+((C/x)̂D))))

A=Y_(Min)

B=Y_(max)

C=EC₅₀

D=Hill Slope

GI₅₀ is the concentration of the compound when Y=(Y_(max)+Yt₀)/2

IC₅₀ is the concentration of the compound when Y=50% of DMSO control

Y=Cell viability measured as luminescence unit

t₀=time when compound was added

Proliferation and apoptosis were measured using CellTiter-Glo andCaspase 3/7-Glo. CalX2 values are the lowest concentration at whichCompound 1 induces a 2-fold increase of cleaved caspase 3/7 compared toDMSO control. Proliferation and apoptosis data is the average of 3experiments.

TABLE 45 Effect of Compound 1 on HCC cell line proliferation. Cell LineGI₅₀ IC₅₀ Cal_X2 JHH-1 0.0016 0.0946 0.0427 JHH-5 0.0045 0.0072 0.0139Hep3B 0.0053 0.0147 0.0028 HuH-7 0.0212 0.4894 0.0118 HuCCT1 0.02531.3033 0.0213 HuH-6-Clone5 0.0291 1.2236 1.5813 SNU-387 0.0332 0.10410.0046 HepG2 0.0346 1.2420 0.0129 SNU-182 0.0764 4.9775 5.2385 JHH-70.0834 0.5476 4.7601 JHH-2 0.1289 4.4850 0.2806 HuH-1 0.2351 7.26436.5641 SNU-398 0.2652 1.9653 0.0378 JHH-4 0.3627 2.3178 0.0588 PLC-PRF-50.8884 4.0089 3.8310 FOCUS 1.4994 4.2962 3.8562 HepG2/C3A 4.6211 10.00000.8273 HLE 4.8451 9.6157 10.0000 SNU-423 6.2355 10.0000 10.0000 HLF6.6814 7.3878 7.2156 SK-HEP-1 7.0390 10.0000 10.0000 SNU-475 9.987910.0000 10.0000 JHH-6 10.0000 10.0000 10.0000 SNU-449 10.0000 10.000010.0000

Conclusion:

Compound 1 inhibits proliferation and induces apoptosis in multiple HCClines.

Anti-Proliferative Activity Across a Panel of 64 Cancer Cell Lines.

Cells were treated with DMSO or increasing concentrations of Compound 1for 72 h. Proliferation was measured using CellTiter-Glo as described.Results are shown in Table 46.

TABLE 46 Anti-proliferative activity of Compound 1 across a panel of 64cancer cell lines. Cell line Tumor Type GI₅₀ (μM) IC₅₀ (μM) SW48 Colon0.0057 0.088 MALME-3M Melanoma 0.0011 0.0038 HT29/219 Colon 0.00170.0045 HCT-116 Colon 0.017 0.022 LOX-IMVI Melanoma 0.022 0.025 HT29Colon 0.016 0.025 A375 Melanoma 0.021 0.024 Colo 205 Colon 0.025 0.040AGS Stomach 0.023 0.028 JHH-5 Liver 0.0045 0.007 SW620 Colon 0.047 0.092MiaPaCa-2 Pancreas 0.047 0.80 JHH-5 Liver 0.0045 0.0072 SW620 Colon0.0474 0.0918 MiaPaCa-2 Pancreas 0.0471 0.0798 JHH-1 Liver 0.0016 0.0946NCI-H2122 Lung 0.0318 0.0427 Hep3B Liver 0.0053 0.0147 NCI-H1755 Lung0.0404 0.0584 92-1 Melanoma 0.0102 0.0316 BxPC-3 Pancreas 0.0368 0.0708SW1417 Colon 0.0005 0.0169 HOP92 Lung 0.1077 0.1173 NCI-H23 Lung 0.03640.1821 PC-9 Lung 0.2167 0.3791 HuH-7 Liver 0.0212 0.4894 MEL-202Melanoma 0.0385 0.0968 SW900 Lung 0.0048 0.0217 NCI-H1299 Lung 0.23360.4982 A549 Lung 0.0402 0.0822 LOVO Colon 0.0630 0.1256 NCI-H460 Lung0.2441 0.6445 SNU-387 Liver 0.0332 0.1041 HuCCT1 Liver 0.0253 1.3033HOP62 Lung 0.3390 3.4861 HuH-6-Clone5 Liver 0.0291 1.2236 JHH-7 Liver0.0834 0.5476 NCI-H838 Lung 0.5670 9.1808 NCI-H226 Lung 1.6266 6.1499NCI-H28 Lung 1.2797 2.3574 MDA-MB-231 Breast 0.0353 3.3333 JHH-2 Liver0.1289 4.4850 HepG2 Liver 0.0346 1.2420 RPMI-8226 Multiple 3.2365 9.7392myeloma K-562 Leukemia 5.4223 6.0279 SNU-182 Liver 0.0764 4.9775 HuH-1Liver 0.2351 7.2643 SNU-398 Liver 0.2652 1.9653 JHH-4 Liver 0.36272.3178 PLC-PRF-5 Liver 0.8884 4.0089 FOCUS Liver 1.4994 4.2962 HepG2/C3ALiver 4.6211 10.0000 HLE Liver 4.8451 9.6157 SNU-423 Liver 6.235510.0000 HLF Liver 6.6814 7.3878 SK-HEP-1 Liver 7.0390 10.0000 SNU-475Liver 9.9879 10.0000 JHH-6 Liver 10.0000 10.0000 SNU-449 Liver 10.000010.0000 NCI-H441 Lung 0.1838 6.3503 NCI-H1703 Lung 1.3513 1.6795NCI-H1975 Lung 2.0476 3.1940 NCI-H520 Lung 5.2445 8.3699 CFPAC-1Pancreas 1.9512 7.3967 PANC-1 Pancreas 5.4360 10.0000 KATOIII Stomach7.0455 8.0240

Compound 1 was shown to inhibit the proliferation of multiple cancercell lines derived from CRC, melanoma, gastric cancer, HCC, lung cancer,pancreatic cancer, leukemia, and multiple myeloma.

Anti-Proliferative and Apoptotic Activity in BRAF Mutant andBeta-Catenin Mutant or Active Cancer Cell Lines.

The mutation status of BRAF, CTNNB1, KRAS, and EGFR in five cell linesevaluated was based on public data (COSMIC and CCLE) and confirmedinternally. β-catenin status was evaluated using TOP Flash reportersystem by transient transfection. A cell line was defined as β-cateninactive if a ratio of Top Flash reporter over Fop Flash reporter isgreater than 2. N/A: Not available. Transfection efficiency in Colo 205(BRAF V600E) was too low to access its β-catenin activity using thisapproach. Antiproliferative and apoptotic activity of Compound 1 in thefive cell lines were measured as described above.

TABLE 47 Antiproliferative and apoptosis activity of Compound 1 in BRAFmutant and beta-catenin mutant and active cell lines. Apoptosis TumorMutation status of β-catenin Proliferation IC₅₀ induction CalX2 Celllines type key genes status (μM) (μM) Colo 205 CRC BRAF (V600E) N/A0.036 +/− 0.023 0.053 +/− 0.039 LOX-IMVI Melanoma BRAF (V600E) Inactive0.025 +/− 0.008 0.034 +/− 0.028 SW48 CRC CTNNB1 (S33Y); Active 0.009 +/−0.007 0.005 +/− 0.001 EGFR (G179S) AGS Gastric CTNNB1 (G43E); Active0.028 +/− 0.021 0.004 +/− 0.002 KRAS (G12D) Hep3B HCC — Active 0.014 +/−0.006 0.002 +/− 0.002

Compound 1 potently inhibits proliferation and induces apoptosis in bothBRAF mutant and beta-catenin mutant or active cancer cell lines,including BRAF mutant CRC, BRAF mutant melanoma, beta-cateninmutant/EGFR mutant CRC (i.e. beta-catenin active/EGFR mutant CRC),beta-catenin mutant/KRAS mutant gastric cancer (i.e. beta-cateninactive/KRAS mutant gastric cancer), and HCC.

Oncogenic Pathway Inhibition. Effect on MAPK Signaling.

Cancer cells were seeded at a density of 25,000 cells per well in96-well tissue culture plates and incubated at 37° C. in a CO₂ incubatorovernight. After treatment with Compound 1 at 37° C. for 2 h, the cellswere lysed with Mesoscale lysis buffer and pRSK S380 levels in eachlysate were measured via Mesoscale ELISA technology.

Conclusion.

Compound 1 potently inhibited pRSK1 in multiple cancer cell lines (Table48).

TABLE 48 Compound 1 pRSK1 S380 IC₅₀ Values in BRAF Mutant LOX-IMVI andColo 205 Cancer Cell Lines. Cell line n = 3 pRSK1 5380 IC₅₀ (μM)LOX-IMVI 0.038 +/− 0.009 Colo 205 0.047 +/− 0.01  SW48 0.021 +/− 0.001AGS 0.020 +/− 0.001

In a time course experiment, Colo-205 cancer cells were treated with 0.5μM Compound 1 for various time periods. The effect of Compound 1 on pRSKS380 was measured as described. The effect of Compound 1 on other MAPKpathway markers (DUSP4 and DUSP6) was measured via Western blotting withspecific antibodies. The time course data in FIG. 188 indicates Compound1 causes sustained inhibition (up to 72 hr) of the following ERKtargets: pRSK1, DUSP4 and DUSP6. BRAF inhibitors (BRAFi) do not causesustained ERK inhibition in BRAF mutant CRC lines (Corcoran et al.,Cancer Discov. 2012, 2:227-35). Sufficient and sustained inhibition ofERK seems to be critical for clinical efficacy of BRAFi and MEKinhibitors (MEKi) in BRAF mutant melanoma (Bollag et al., Nat Rev DrugDisc. 2012; 11, 873-886) and CRC patients (Corcoran et al., CancerDiscov. 2012, 2:227-35). Lack of sustained inhibition of ERK by BRAFimay contribute to the lack of clinical activity of BRAFi in BRAF mutantCRC patients. The sustained inhibition of ERK by Compound 1 may providean advantage over BRAFi in BRAF mutant CRC patients.

The ability of Compound 1 to inhibit MAPK signaling was assessed bydetermining the DUSP4 and DUSP6 protein expression. Colon cancer cellline Colo 205 (BRAF V600E) cultures were treated with DMSO or increasingconcentrations of Compound 1 for 2, 8 or 24 h. Proteins were extractedfrom treated cells and analyzed by Western blot using antibodies againstDUSP4, DUSP6, cyclin D1, c-Myc, YAP or β-actin. RNAs were extractedusing Cell-To-CT kit and quantitative PCR was performed with probesspecific for DUSP4, DUSP6, SPRY2, c-Myc and cyclin D1. Specific probesfor β-actin were used for normalization.

In Colo 205 (BRAF V600E), DUSP4 and DUSP6 were significantly reduced byCompound 1 as early as 2 h and the reduction was sustained through 24 h(FIG. 189A). Compound 1 treatment led to the reduction of SPRY2transcription in a concentration-dependent manner in Colo 205 (FIG.189B), consistent with potent ERK inhibition. Levels of cyclin D1 andc-Myc, which are downstream of both canonical Wnt and MAPK signaling,were assessed. Compound 1 significantly decreased cyclin D1 and c-MycRNA and protein levels in Colo 205 cells (FIGS. 189A-C). Compound 1treatment resulted in decreased YAP protein at 24 h in Colo 205 (FIG.189A). Taken together, our cellular data is consistent with strong,sustained MAPK pathway inhibition.

To further evaluate the ability of Compound 1 to inhibit MAPK signaling,RNA expression was assessed of additional MAPK targets (BMF, DUSP5,DUSP6, EFNA1, EGR1, ETV5, FOS, FOSL1, GJA1, IL-8, SPRY2, and SPRY4).Cultures of the colon cancer cell lines Colo 205 (characterized by aBRAF V600E mutation) and HT-29 (characterized by a BRAF V600E mutation)were treated with DMSO or Compound 1 at 0.3 or 1 μM for 6 h. RNAs wereextracted using MagMAX Total RNA Isolation kit and quantitative PCR wasperformed with probes specific for BMF, DUSP5, DUSP6, EFNA1, EGR1, ETV5,FOS, FOSL1, GJA1, IL-8, SPRY2, SPRY4. Specific probes for 18S rRNA wereused for normalization.

In both cell lines, mRNA levels of DUSP5, DUSP6, EGR1, ETV5, FOS, FOSL1,IL-8, SPRY2, SPRY4 were reduced by Compound 1 (FIGS. 189D-I), consistentwith ERK inhibition. The finding that mRNA levels of GJA1 are reduced inColo205 cells and increased in HT29 may be related to our finding thatCompound 1 is cytotoxic in Colo205 and cytostatic in HT29. Compound 1treatment resulted in increased mRNA levels of BMF and EFNA1 at 6 h inColo 205 and HT-29. Taken together, our cellular data is consistent withMAPK pathway inhibition.

Effect on Beta-Catenin and YAP Signaling.

Cellular activity against beta-catenin and YAP target genes by Compound1 was evaluated. Colon cancer cell line Colo 205 (BRAF V600E) cultureswere treated with DMSO or increasing concentrations of Compound 1 for 2,8 or 24 h. RNAs were extracted using Cell-To-CT kit and quantitative PCRwas performed with probes specific for Axin2, CTGF, and AREG. Specificprobes for β-actin were used for normalization.

Compound 1 treatment led to increased Axin2 RNA (FIG. 190A). Compound 1significantly reduced the expression of Hippo/YAP target genes (CTGF,AREG) in Colo 205 (BRAF V600E) at 2, 8 and 24 hr (FIG. 190A). Takentogether, these data suggest that Compound 1 impacts Wnt signaling andblocks Hippo signaling in Colo 205 cancer cells.

Cellular activity against additional YAP target genes by Compound 1 wasevaluated (FIG. 190B-E). Cultures of the colon cancer cell lines Colo205 and HT-29 were treated with DMSO or Compound 1 at 0.3 or 1 μM for 6h. RNAs were extracted using MagMAX Total RNA Isolation kit andquantitative PCR was performed with probes specific for CYR61, CITED2,CXCL1, ELF3, HAS2, HES1, and MAFF. Specific probes for 18S rRNA wereused for normalization.

In both cell lines, mRNA levels of CYR61, CXCL1, HAS2, HES1 and MAFFwere reduced by Compound 1. The finding that CYR61 mRNA levels arereduced in Colo205 cells but not in HT29 and that mRNA levels of CITED2are increased in HT29, but not in Colo205, may be related to our findingthat Compound 1 is cytotoxic in Colo205 and cytostatic in HT29. Compound1 treatment resulted in increased mRNA levels for CITED2 and ELF3 mRNAat 6 h in Colo 205 and HT-29. (FIG. 190B) Taken together, our cellulardata is consistent with YAP pathway inhibition.

Evaluation of Sensitivity in Cell Lines Having Beta-Catenin Mutations.

The effect of Compound 1 on cell lines having β-catenin mutations wasevaluated. (FIG. 205 and FIGS. 206A-B). Compound 1 showed efficacyagainst cell lines with mutated β-catenin. Such cell lines demonstratethat cancers characterized by mutated β-catenin are more sensitive totreatment with Compound 1. Compound 1 was further shown to modulateβ-catenin, and YAP in BRAF and CTNNB1 mutant cell lines as shown in FIG.207. Compound 1 also modulates target gene expression controlled byMAPK, β-catenin, and YAP in BRAF and CTNNB1 mutant cell lines asprovided in FIGS. 208A-B.

Western Blot.

Compound 1 modulation of MAPK, WNT/β-catenin, and Hippo/YAP pathwaymarkers was evaluated by standard Western blotting. LOX-IMVI, SW48, andColo-205 cells were plated in 6-well plates at a density of 250,000cells per well and were allowed to attach overnight. Compound 1 wasadded to cells at concentrations of 0.03, 0.1, 0.3, 1, and 3 μM fordurations of 2, 8, and 24 hours. Cells were harvested and lysed in RIPAbuffer (50 mM Tris-HCl, pH 7.4, 150 mM sodium chloride [NaCl], 0.25%deoxycholic acid, 1% Nonidet P-40, 1 mM ethylenediaminetetraacetic acid[EDTA], protease and phosphatase inhibitors). The cell lysates wereheated in sodium dodecyl sulfate (SDS)-sample buffer and 40 μg of celllysate per condition were loaded onto gels and separated using SDSpolyacrylamide gel electrophoresis (PAGE). Protein was transferred tonitrocellulose membrane, and immunoblotted with anti DUSP4, DUSP6, cMyc,Cyclin D1, YAP, AXIN2, HDACS (phospho S498), and β-actin antibodies.Membranes were scanned on the Licor Odyssey system.

Quantitative Polymerase Chain Reaction.

Compound 1 modulation of MAPK, WNT/β-catenin, and Hippo/YAP pathwaygenes was evaluated by real-time (RT)-qPCR. Lysyl oxidase IMVI, SW48,and Colo-205 cells were plated in 96-well plates at a density of 20,000cells per well and were allowed to attach overnight. Compound 1 wasadded to cells at half log concentrations from 1 nM to 10 μM fordurations of 2, 8, and 24 hours. Cells were harvested using the TaqManGene Expression Cells-to-CT Kit according to the product manual. Next,RT-PCR was performed and the resulting cDNA was used in qPCR reactionson the ViiA7 Real-Time PCR System (Thermo Fisher Scientific). TaqManprobes were used to monitor changes in DUSP4, DUSP6, SPRY2, MYC, CCND1,AXIN2, CTGF, Cyr61, AREG, and ACTB genes. All genes were normalized toACTB expression and reported as percentage of DMSO-only control.

Gene Expression Analysis:

Human bronchial epithelial cells were cultured in T-150 flasks in BEpiCMgrowth medium and allowed to reach 80% confluency. Cells were plated in12-well plastic culture plates at 150,000 cells per well in BEpiCMmedium for 24 hours. After a 24-hour incubation, cells were treated withdimethyl sulfoxide (DMSO) as a control, Compound 1 at 0.1, 1, 10 μM, for30 minutes. Cells were then stimulated with 100 ng/ml recombinant Wnt3a(formulated in phosphate buffered saline [PBS]), 350 pM RSPO3(formulated in PBS) or a combination of Wnt3 and RSPO3 for 24 hours.Ribonucleic acid (RNA) was isolated using a Qiagen Rneasy Mini Kitaccording to manufacturer's instruction. Axin2 and gene expression wasdetermined using reverse transcription polymerase chain reaction(RT-PCR) Taq-Man assays. Quantitative PCR (qPCR) was performed usingSuperScript® III One-Step RT-PCR System and ran on a Viia 7 Real-TimePCR System. Data was normalized to glyceraldehyde 3-phosphatedehydrogenase. Compound 1 inhibits Axin2 expression in human bronchialepithelial cells. Gene expression was measured at 24 hours. From theseresults it was shown that Compound 1 inhibits Axin2 expression in humanbronchial epithelial cells. (FIG. 209).

Long Term Colony Assay.

Compound 1 was assessed for its ability to inhibit the colony formationof cancer cells via a long-term colony forming assay. Cells andcompounds were added to 96-well plates and were monitored for up to 8weeks for the formation of colonies. Compound and media were replenishedevery 1 week throughout the course of the assay. Colony formation wasdetected via imaging at 4× on the IncuCyte ZOOM System. Compound 1demonstrated inhibition of colony formation of β-catenin mutant cells ata level greater than MEK inhibitors (trametinib) and ERK inhibitors(GDC0994). SW48 (colo) cells, HCT-116 (colo) cells, AGS (gastric) cells,and Hep3B (HCC) cells were treated with Compound 1 and showed greaterlevels of inhibition than seen with treatment with MEK inhibitors or ERKinhibitors. (FIGS. 210A-210D). Compound 1 was further shown tosurprisingly inhibit colony formation of AGS cells that are resistant toMEK inhibitor treatment with trametinib. Such results suggest Compound 1can be useful in treating cancers resistant to other treatments.

Evaluation of Immunomodulatory Effects.

The effect of Compound 1 was evaluated on PD-L1 expression levels. Cellswere cultured in presence or absence of Compound 1 for indicated timebefore expression levels of PD-L1, DUSP4 and α-tubulin or α-actin weremeasured by Western blot. To detect surface levels of PD-L1, cells weretreated with DMSO or Compound 1 at indicated concentrations for 48 h andcell surface expression of PD-L1 was detected using flow cytometryanalysis (FACS) with an APC-labeled antibody to PD-L1 (clone 29E.1A3;BioLegend, San Diego, Calif.). Geometric mean of PD-L1 positive cellswas determined by FlowJo 10 (Treestar, Ashland, Oreg.).

Conclusion.

Compound 1 directly inhibits PD-L1 expression in multiple cancer cellsincluding HOP62, KARPAS-299, and LOX-IMVI (BRAF V600E) (FIG. 191A). FACSanalysis indicates that surface PD-L1 levels are also inhibited byCompound 1 in multiple cancer cell lines (FIG. 191B).

To determine if Compound 1 down-regulation of PD-L1 enhances T cellactivation, compound-treated KARPAS-299 cancer cells were co-culturedwith PBMC-derived T cells stimulated with low concentrations of superantigen (SEB). KARPAS-299 cells were treated with DMSO (D) or Compound 1at indicated concentrations for 48 h. PBMC from healthy donors weretreated with or without 20 ng/ml SEB for 48 h. After wash with PBS, thePBMCs were incubated with the cancer cells for 24 h and the supernatantswere collected to measure IL-2 and IFNγ using Mesoscale assays.

Supernatant levels of IL-2 and IFNγ were used as functional markers of Tcell activation. In the absence of SEB, PBMC co-cultured withCompound-1-treated KARPAS-299 cells produced little IL-2 or IFNγ. In thepresence of low concentrations of SEB (20 ng/ml), Compound 1-treatedcancer cells co-cultured with PBMC demonstrated increased levels of bothIL-2 and IFNγ production (FIGS. 192A-B). The increased levels of IL-2and IFNγ in Compound 1-treated cancer cells were similar to the levelsobserved with treatment of anti-PD-L1 (Ultra-LEAF™ from Biolegend).

The effect of Compound 1 treatment on levels of IL-8 was determined inPBMC culture media. PBMCs were isolated from whole blood and cultured inRPMI media plus 10% FBS. PBMCs were plated at 1×10⁶ per milliliter in 10cm² dishes. The PBMCs were treated with 0.1% DMSO or 0.5 μM Compound 1.Treatments were taken down at the designated time points. The culturemedia (1 mL) was used for IL-8 analysis. The IL-8 analysis was performedwith a Mesoscale V-Plex Human IL-8 kit according to the manufacturer'sinstructions. Compound 1 was shown to inhibit IL-8 levels at differenttime-points (FIG. 192C).

TEAD Reporter Assay.

TEAD reporter activity was analyzed using WI38 VA13 cells stablyexpressing a YAP/TAZ responsive synthetic promoter driving luciferaseexpression (8×GTIIC-luciferase). 10,000 cells per well were seeded on awhite-walled 96-well plate and left overnight. After 16-20 hours, cellswere treated with compound and TEAD reporter activity was measured 24 or72 hours later using Bright Glo luciferase assay (Promega) according tothe manufacturer's instructions. This assay was performed 3 times forCompound 1 and twice for Trametinib. See FIG. 212.

Viability Assay.

In parallel 10,000 WI38 VA13 cells expressing 8×GTIIC-luciferase wereseeded in each well of a black-walled 96-well plate. After 16-20 hourscells were treated with compound for 24 or 72 hours. At this time theserum and compound containing media was removed and replaced with 100 μlserum free media and 100 μl Cell Titer Fluor (Promega). The plate wasincubated for 2 hours at 37° C. before reading fluorescence output. Thisassay is based on measurement of live-cell protease activity. Theviability assay was performed to confirm that any effects of compoundson TEAD reporter were not the result of compound effects on viability.This assay was performed 3 times for Compound 1 and twice forTrametinib.

Conclusion.

These data provide an additional therapeutic hypothesis suggesting thattreatment with Compound 1 will potentiate T cell activation. The invitro data suggests that Compound 1 may enhance T cell immunity againstcancer cells by inhibiting key oncogenic pathways such as the MAPKpathway and down-regulating the immune checkpoint molecule PD-L1expression in tumor microenvironment. Cancer types that express highlevels of PD-L1 (for example, melanoma, lung, RCC, or HCC) may thereforebe sensitive to Compound 1.

Animal Models

Xenograft Models.

For xenograft model studies human cancer cell lines were injected intoSCID (severe combined immunodeficiency) mice. Cancer cell lines werepropagated in culture in vitro. Tumor bearing animals were generated byinjecting precisely determined numbers of cells into mice. Followinginoculation of animals, the tumors were allowed to grow to a certainsize prior to randomization. The mice bearing xenograft tumors rangingbetween pre-determined sizes were pooled together and randomized intovarious treatment groups. A typical efficacy study design involvedadministering one or more compounds at various dose levels totumor-bearing mice. Additionally, reference chemotherapeutic agents(positive control) and negative controls were similarly administered andmaintained. Tumor measurements and body weights were taken over thecourse of the study.

Mice were anesthetized with inhaled isoflurane and then inoculated withLOX-IMVI tumor cells subcutaneously above the right hind leg with 0.1 mLof a single cell suspension in PBS using a sterile 1 mL syringe fittedwith a 26-gauge needle. Following inoculation of the animals, tumorswere allowed to grow to approximately 75-125 mm³ or in some cases250-400 mm³ prior to randomization of the mice. The tumor of each animalwas measured and animals with tumors in the appropriate range wereincluded in the study. Animals from the study pool were then distributedrandomly into various cages and the cages were randomly assigned tovehicle, positive control, or test article groups. All of the mice weretagged with metal ear tags on the right ear. A typical group consistedof 8-10 animals. For a typical xenograft study, SCID mice bearing tumorswere randomized and dosed with compounds ranging from, for example, 100mg/kg to 0.1 mg/kg with different dose scheduling, including, but notlimited to, qd, q2d, q3d, q5d, q7d and bid. The mice were dosed for 1-4weeks. Tumors were measured twice a week using calipers and tumorvolumes were calculated using the formula of W²×L/2.

The purpose of these studies was to test the efficacy of Compound 1 inthe cell line-derived xenograft models, LOX-IMVI (melanoma) and Colo205(colorectal) and the PDX1994060146 (patient-derived xenograft [PDX146])colorectal xenograft model. These models were chosen because they harborthe V600E BRAF mutation. Additional PK/PD analysis was performed toexamine the Compound 1-mediated inhibition of pathway biomarkers in thePDX146 xenograft model.

LOX-IMVI Subcutaneous Melanoma Xenograft Model.

The purpose of this study was to confirm the efficacy of Compound 1 inthe LOX-IMVI melanoma xenograft model. One study (FIG. 193) in theLOX-IMVI xenograft model testing two dose levels of Compound 1 (15 and30 mg/kg) demonstrated significant tumor volume reduction compared tothe vehicle control (p<0.001 for both dose levels). Tumor regression wasobserved in 9 out of 9 animals for both dose levels and 1 out of 9animals from each group was tumor free at study end.

In a separate experiment, Compound 1 was administered orally, QD for 8days at 0.2, 1, 5, 10, and 15 mg/kg. Dose-dependent antitumor activitywas observed with Compound 1 treatment in the LOX-IMVI xenograft model(FIG. 194). Tumor regression was observed at the 10 and 15 mg/kg doselevels.

Colo 205 Subcutaneous Colorectal Xenograft Model. Colo 205 SubcutaneousColorectal Xenograft Model.

The purpose of these studies was to test the efficacy of Compound 1 inthe Colo 205 colorectal cancer xenograft model, and determine whethertwice daily dosing (BID) had an impact on antitumor activity. In thefirst experiment Compound 1 was administered orally, QD for 15 days at0.2, 1, 5, 10, and 15 mg/kg. Dose-dependent antitumor activity wasobserved with Compound 1 treatment in the Colo 205 xenograft model (FIG.195). A scheduling study was conducted to determine whether BID dosingincreased the antitumor activity of Compound 1. Dose-dependent antitumoractivity was observed with Compound 1 treatment in the Colo 205xenograft model (FIG. 196).

PDX1994060146 Subcutaneous Colorectal Patient-Derived Xenograft Model.

The purpose of these studies was to test the efficacy of Compound 1 inthe PDX1994060146 (PDX146) colorectal cancer xenograft model anddetermine whether BID dosing had an impact on antitumor activity. A timeto progression (TTP) study was performed to determine the effect oflonger treatment duration on tumor growth.

In the first experiment Compound 1 was administered orally, QD at 1, 5,and 15 mg/kg or 5 and 15 mg/kg BID for 22 days. Dose-dependent antitumoractivity was observed with Compound 1 treatment in the PDX146 xenograftmodel (FIGS. 197A-B). Dosing 15 mg/kg BID appeared to increase theantitumor activity of Compound 1 compared to the administration of 15mg/kg QD.

In the TTP study, Compound 1 was administered orally, 1, 5, and 15 mg/kgBID for 49-77 days. Compound 1 treatment groups were dosed throughoutthe duration of the study until the group mean reached the predeterminedendpoint of approximately 1200 mm³ or study termination. Tumor growthdelay (TGD) was calculated as the time between the termination of thevehicle control group (on day 43) and the Compound 1 treatment groups.The TGD was 8, 12 and >37 days for the 1, 5 and 15 mg/kg treatmentgroups, respectively. (FIG. 198)

Biomarkers representing the activity of three different pathways, MAPK,Wnt, and Hippo, were inhibited in the PDX146 xenograft model. Sustainedinhibition of these pathway biomarkers was observed through 24 h.

Antitumor Activity of Compound 1 in the β-catenin Mutant SW48 ColorectalXenograft Model.

Female SCID mice were inoculated with 2×10⁶ SW48 tumor cells into theright flank. Mice were randomized into treatment groups (n=10/group) atthe time of treatment initiation. Test article treatment started on Day10 when the tumors were approximately 110 and 105 mm³. (See FIGS.202A-B.) Black dotted line is the tumor volume at the initiation ofdosing. Graph on the left is a dose-response study. Graph on the rightis a time to progression study where animals were maintained on drugduring the course of the study. Dotted line is the tumor volume on Day28 when the vehicle control group was terminated.

Antitumor Activity in the Orthotopic Hep3B2.1-7 Hepatocellular CarcinomaXenograft.

Female SCID mice were orthotopically inoculated with 2×10⁶ Hep3B2.1-7tumor cells per animal. Seven days post-inoculation the animals wererandomized into treatment groups based on body weight and the treatmentcommenced (Study day 0). Take rate assessment of a satellite groupconfirmed the presence of tumor in the liver in 100% of the animals.Treatment with Compound 1 was started and Compound 1 was dosed orally,QD for 21 days. Significant mean body weight loss expected with thismodel was observed in the vehicle control group. Animals treated with 15mg/kg Compound 1 showed minimal body weight loss and a significant meanbody weight gain was observed in the 30 mg/kg Compound 1 treatmentgroup. On the day of study termination, the tumors were removed andweighed. Individual tumor weights and the mean tumor weight ±SEM of eachgroup was plotted (FIG. 203). Percent inhibition was calculated relativeto the vehicle control. P values were derived from a one-way ANOVA witha Dunnet's post-hoc analysis. ***=p<0.001.

Antitumor Activity of Compound 1 in the C-Met Amplified HepatocellularCarcinoma Patient-derived Xenograft Model, L10612.

Female SCID mice were inoculated with hepatocellular carcinoma PDX modelLI0612 tumor fragments (2-4 mm in diameter) into the right flank. Themice were randomized into treatment groups (n=10/group) at the time oftreatment initiation. Test article treatment started on Day 18 when thetumors were approximately 150 mm³ in size. Tumor growth progressed inthe vehicle control and Compound 1 treatment groups over the dosingperiod. A change in the growth kinetics was noted with Compound 1administration resulting in significant tumor growth inhibition (TGI)with 30 mg/kg treatment (p=0.038, compared to the vehicle control). SeeFIG. 204.

Pharmacokinetic/Pharmacodynamic Data in a BRAF Mutant Patient-DerivedXenograft Model.

Based on the known kinases (ERK 1/2, NLK and SIK) that are inhibited byCompound 1, the impact of compound treatment was evaluated on MAPK,β-catenin and Hippo pathway biomarkers in PDX146 tumors from xenograftedmice. Tumor-bearing mice (tumors were ˜400 mm³) were treated with asingle dose of 1 or 5 mg/kg Compound 1. Tumor tissue was collected at 1,2, 4, 8, and 24 h post-dose.

The modulation of the MAPK pathway was evaluated by examination of tumorDUSP4, DUSP6 and Sprouty (SPRY2) mRNA levels and pRSK and pERK proteinlevels. DUSP6 mRNA levels were significantly decreased with compoundtreatment starting 2 hr post-dose and remained suppressed through 24 hat both dose levels (FIG. 199A). A similar pattern was observed withDUSP4 and SPRY2 mRNA levels (FIGS. 200A-B). Phospho-RSK (pRSK) andphospho-ERK (pERK) protein levels were modulated by Compound 1 treatmentin a dose- and time-dependent manner (FIGS. 201A-D). Levels of cMyc(FIG. 199B) and cyclin D1 (FIG. 200C), which are downstream of both theMAPK and Wnt signaling pathways, were inhibited with Compound 1treatment. Compound 1 treatment upregulated the Wnt target gene, Axin2.Treatment with Compound 1 at both dose levels demonstrated a significantincrease in Axin2 mRNA levels 24 h post-dose. Sustained inhibition ofAREG (a downstream target gene in the Hippo pathway) mRNA levels wasobserved through 24 h. Additionally Compound 1 inhibited YAP proteinlevels in a time-dependent manner (not statistically significant (seeFIG. 200D), which could be due to SIK inhibition and Hippo pathwayregulation or an indirect effect as a result of MAPK inhibition.

These data suggest that Compound 1 impacts three different pathways,MAPK, Wnt and Hippo, in this BRAF mutant colorectal PDX model followinga single dose administration.

Other Efficacy Model Data:

Compound 1 was profiled in additional xenograft models includingβ-catenin mutant (SW48, colorectal) and β-catenin activated models(orthotopic Hep3B, hepatocellular) and a c-met-amplified hepatocellularPDX model (LI0612). Significant antitumor activity was observed in allmodels.

Conclusion:

Significant dose-dependent antitumor activity was observed in all threeBRAF mutant xenograft models (See FIGS. 202A-B, FIG. 203, and FIG. 204).Tumor regression was observed with Compound 1 treatment across themodels and there was a significant growth delay with long term treatmentin the PDX146 model.

Patient Enrichment and Tumor Indications.

Based upon the in vitro and in vivo data of Compound 1, the patientenrichment hypotheses and tumor indications are outlined in Table 49 andTable 50.

TABLE 49 Patient enrichment biomarkers and tumor indications. PatientEnrichment Biomarkers Tumor indications BRAF mutant CRC, Thyroid,Melanoma, Lung NRAS mutant Melanoma KRAS mutant Lung, CRC, PancreasCTNNB1 (β-catenin CRC, Stomach, HCC, Sarcoma mutant and/or active)

TABLE 50 Molecular Alterations Pathways Clinical Indications CTNNB1mutant, Wnt/b-catenin// Hippo HCC YAP amplification BRAF mutant,MAPK//Wnt/b-catenin CRC CTNNB1 CTNNB1 mutant Wnt/b-catenin Gastric BRAFmutant, NRAS MAPK Melanoma mutant

A number of references have been cited, the disclosures of which areincorporated herein by reference in their entirety.

Although the invention has been described with reference to thedisclosed embodiments, those skilled in the art will readily appreciatethat the specific examples and studies detailed above are onlyillustrative of the invention. It should be understood that variousmodifications can be made without departing from the spirit of theinvention. Accordingly, the invention is limited only by the followingclaims.

1. A crystal form comprising Compound 1, or a tautomer thereof:

which has an X-ray powder diffraction pattern comprising peaks at 7.3,8.5, 18.2, and 21.3° 2θ (±0.2° 2θ). 2.-9. (canceled)
 10. A crystal formcomprising a citrate salt of Compound 1, or a tautomer thereof:

which has an X-ray powder diffraction pattern comprising peaks at 4.8,9.6, 18.9, and 19.2° 2θ (±0.2° 2θ). 11.-12. (canceled)
 13. A crystalform comprising a hydrochloride salt of Compound 1, or a tautomerthereof:

which has an X-ray powder diffraction pattern comprising peaks at 9.0,9.8, 9.9, and 16.7° 2θ (±0.2° 2θ). 14.-20. (canceled)
 21. Apharmaceutical composition comprising said solid form of claim 1, and apharmaceutical carrier.
 22. A method for treating or preventing acancer, comprising administering to a subject in need thereof aneffective amount a solid form of claim 1, wherein the cancer is a solidtumor or a hematological cancer.
 23. The method of claim 22, wherein thesolid tumor is melanoma, colorectal cancer, stomach cancer, head andneck cancer, thyroid cancer, bladder cancer, CNS cancer, lung cancer,pancreatic cancer, or soft tissue cancer.
 24. The method of claim 22,wherein the cancer is bladder cancer, breast cancer, CNS cancer, coloncancer, gastrointestinal cancer, endocrine cancer, female genitourealcancer, head and neck cancer, hematopoietic cancer, kidney cancer, livercancer, lung cancer, melanoma, pancreas cancer, prostate cancer, or softtissue cancer.
 25. The method of claim 22, wherein the cancer is glioma,neuroblastoma, stomach cancer, thyroid cancer, adrenal gland cancer,cancer of the uterus, cervix, ovary clear cell, or vulva, leukemia,myeloma, NSCLC, SCLC, sarcoma or osteosarcoma.
 26. A method for treatingor preventing hepatocellular carcinoma (HCC), comprising administeringto a subject in need thereof an effective amount a solid form ofclaim
 1. 27. A method for treating or preventing colorectal cancer(CRC), melanoma, gastric cancer, HCC, lung cancer, pancreatic cancer,leukemia, or multiple myeloma), comprising administering to a subject inneed thereof an effective amount a solid form of claim
 1. 28. A methodfor treating or preventing a cancer expressing PD-L1, comprisingadministering to a subject in need thereof an effective amount a solidform of claim
 1. 29. The method of claim 28, wherein the PD-L1expressing cancer is melanoma, lung cancer, renal cell carcinoma (RCC),or HCC.
 30. A method for treating or preventing a cancer characterizedby a BRAF mutation, comprising administering to a subject in needthereof an effective amount a solid form of claim
 1. 31. The method ofclaim 30, wherein the cancer characterized by a BRAF mutation is CRC,thyroid cancer, melanoma or lung cancer.
 32. (canceled)
 33. A method fortreating or preventing a cancer characterized by an NRAS mutation,comprising administering to a subject in need thereof an effectiveamount a solid form of claim
 1. 34. The method of claim 33, wherein thecancer characterized by an NRAS mutation is melanoma.
 35. A method fortreating or preventing a cancer characterized by a KRAS mutation,comprising administering to a subject in need thereof an effectiveamount a solid form of claim
 1. 36. The method of claim 35, wherein thecancer characterized by a KRAS mutation is CRC, pancreas cancer or lungcancer.
 37. A method for treating or preventing a cancer characterizedby a beta-catenin mutation, comprising administering to a subject inneed thereof an effective amount a solid form of claim
 1. 38. The methodof claim 37, wherein the beta-catenin mutation is one or more ofbeta-catenin S33Y, G34E, S45del, or S33C.
 39. The method of claim 37 or38, wherein the HCC is characterized by a beta-catenin mutation and/orincreased YAP expression.
 40. The method of claim 37, further comprisingan EGFR mutation or increased EGFR activity.
 41. (canceled)
 42. Themethod of claim 37, further comprising a BRAF mutation. 43.-45.(canceled)
 46. The method of claim 37, wherein the cancer characterizedby a beta-catenin mutation is CRC, stomach cancer, HCC or sarcoma.
 47. Amethod for treating or preventing a cancer characterized by an activatedbeta-catenin pathway, comprising administering to a subject in needthereof an effective amount a solid form of claim
 1. 48. The method ofclaim 47, wherein the cancer is CRC, stomach cancer, HCC or sarcoma. 49.(canceled)
 50. A method for treating or preventing gastric cancer,comprising administering to a subject in need thereof an effectiveamount a solid form of claim
 1. 51.-52. (canceled)
 53. The method ofclaim 50, wherein the gastric cancer is characterized by a beta-cateninmutation, or by an activated beta-catenin pathway.
 54. (canceled)
 55. Amethod for treating or preventing melanoma, comprising administering toa subject in need thereof an effective amount a solid form of claim 1.56. The method of claim 55, wherein the melanoma is characterized by aBRAF mutation and/or NRAS mutation.
 57. A method for modulating thelevels of a biomarker in a subject having a cancer, comprisingadministering to said subject an effective amount of a solid form ofclaim
 1. 58. (canceled)
 59. The method of claim 57, wherein thebiomarker is ERK, RSK1, DUSP4, DUSP5, DUSP6, BMF, EFNA1, EGR1, ETV5,FOS, FOSL1, GJA1, IL-8, cMyc, Cyclin D1, YAP, SPRY2, SPRY4, Axin2, CTGF,AREG, CYR61, CXCL1, HAS2, HES1, MAFF, CITED2, ELF3, or PD-L1.
 60. Themethod of claim 57, wherein the modulation is measured by measurement ofthe reduction of phosphorylation levels of one or more of ERK and RSK1.61.-62. (canceled)
 63. The method of claim 22, wherein the solid form ofCompound 1 is administered as a pharmaceutical composition comprising atleast one pharmaceutically acceptable excipient.